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2.  Petroleum Ether Extract of Cissus Quadrangularis (Linn.) Enhances Bone Marrow Mesenchymal Stem Cell Proliferation and Facilitates Osteoblastogenesis 
Clinics (Sao Paulo, Brazil)  2009;64(10):993-998.
To evaluate the effects of the petroleum ether extract of Cissus quadrangularis on the proliferation rate of bone marrow mesenchymal stem cells, the differentiation of marrow mesenchymal stem cells into osteoblasts (osteoblastogenesis) and extracellular matrix calcification. This study also aimed to determine the additive effect of osteogenic media and Cissus quadrangularis on proliferation, differentiation and calcification.
MSCs were cultured in media with or without Cissus quadrangularis for 4 weeks and were then stained for alkaline phosphatase. Extracellular matrix calcification was confirmed by Von Kossa staining. marrow mesenchymal stem cells cultures in control media and osteogenic media supplemented with Cissus quadrangularis extract (100, 200, 300 μg/mL) were also subjected to a cell proliferation assay (MTT).
Treatment with 100, 200 or 300 μg/mL petroleum ether extract of Cissus quadrangularis enhanced the differentiation of marrow mesenchymal stem cells into ALP-positive osteoblasts and increased extracellular matrix calcification. Treatment with 300 μg/mL petroleum ether extract of Cissus quadrangularis also enhanced the proliferation rate of the marrow mesenchymal stem cells. Cells grown in osteogenic media containing Cissus quadrangularis exhibited higher proliferation, differentiation and calcification rates than did control cells.
The results suggest that Cissus quadrangularis stimulates osteoblastogenesis and can be used as preventive/ alternative natural medicine for bone diseases such as osteoporosis.
PMCID: PMC2763075  PMID: 19841707
Cissus quadrangularis; Mesenchymal stem cells; Osteoblastogenesis; Osteoporosis; Osteoblast
3.  Basti: Does the equipment and method of administration matter? 
Basti is one of the five procedures of panchakarma in Ayurveda. Classically, it is advocated in the diseases of vata. It is mainly of two types viz. asthapana and anuvasana. According to the classical texts basti administration is done with the help of animal bladder (bastiputaka) and specially prepared metal/wooden nozzle/catheter (bastinetra), the whole assembly is called as bastiyantra. Nowadays, except in some of the Vaidya traditions in Kerala, basti administration is often done using enema-can or douche-set. In the aforesaid classical procedure active pressure is expected to be given on the bastiputaka whereas, in conventionally used enema-can only passive or gravitational force plays a role. This is important in the context of ‘basti danakala or pidanakala i.e. time for basti administration′.
PMCID: PMC3667437  PMID: 23741155
Basti; basti pidanakala; bastiputaka; colon; enema-can
4.  A comparative study of Agni karma with Lauha, Tamra and Panchadhatu Shalakas in Gridhrasi (Sciatica) 
Ayu  2010;31(2):240-244.
Sushruta has mentioned different methods of management of diseases, such as Bheshaja karma, Kshara Karma, Agni karma, Shastrakarma and Raktamokshana. The approach of Agni karma has been mentioned in the context of diseases like Arsha, Arbuda, Bhagandar, Sira, Snayu, Asthi, Sandhigata Vata Vikaras and Gridhrasi. Gridhrasi is seen as a panic condition in the society as it is one of the burning problems, especially in the life of daily laborers. It is characterized by distinct pain starting from Sphik Pradesha (gluteal region) and goes down toward the Parshni Pratyanguli (foot region) of the affected side of leg. On the basis of symptomatology, Gridhrasi may be simulated with the disease sciatica in modern parlance. In modern medicine, the disease sciatica is managed only with potent analgesics or some sort of surgical interventions which have their own limitations and adverse effects, whereas in Ayurveda, various treatment modalities like Siravedha, Agni karma, Basti Chikitsa and palliative medicines are used successfully. Among these, Agni karma procedure seems to be more effective by providing timely relief. Shalakas for Agni karma, made up of different Dhatus like gold, silver, copper, iron, etc. for different stages of the disease conditions, have been proposed. In the present work, a comparative study of Agni karma by using iron, copper and previously studied Panchadhatu Shalaka in Gridhrashi has been conducted. A total of 22 patients were treated in three groups. Result of the entire study showed that Agni karma by Panchadhatu Shalaka provided better result in combating the symptoms, especially Ruka and Tandra, while Lauhadhatu Shalaka gave better results in combating symptoms of Spanadana and Gaurava. In the meantime, Tamradhatu Shalaka provided better effect in controlling symptoms like Toda, Stambha and Aruchi. Fifty percent patients in Panchadhatu Shalaka (Group A) were completely relieved. In Lauhadhatu Shalaka (Group B), the success rate was 00.00%, and in Tamradhatu Shalaka (Group C), the percentage of success rate was 14.28%. After analyzing the data, Tamradhatu Shalaka was found to be more effective than Lauha and Panchadhatu Shalakas.
PMCID: PMC3215372  PMID: 22131718
Agni karma; Gridhrasi; Panchadhatu Shalaka; Parshni Pratyanguli; Sciatica
5.  Study of Preparation and Standardization of ‘Maadhutailika Basti’ with special reference to Emulsion Stability 
Ayu  2010;31(1):1-6.
‘Basti’ is compared with conventional enema due to its similarity in procedure of administration. But in effect enema is a simple evacuation technique, whereas ‘Basti’ is a therapeutic measure considered as ‘Ardha Chikitsa’ i.e half therapy in the disease management. Constitution of ‘Basti dravya’ formulation specifically ‘Aasthaapana Basti’ has a direct impact on the end result of ‘Basti’ therapy. A systematic stepwise procedure of preparation of ‘Aasthaapana Basti’ is described in Ayurvedic Classics. The present study evaluates the significance of this procedure through standardization of ‘Maadhutailika Basti’, a type and standard of ‘Aasthaapana Basti’. Four samples of ‘Aasthaapana Basti’ including the classical one were prepared for this study by replacement, exclusion and supplementation of honey in the formulation. A comparative study of physico-chemical characteristics of these samples was carried out. The stability of the constitution of ‘Maadhutailika Basti’ was also studied to assess the role of honey as a natural emulsifying agent. The study underlines the significance of the procedure described in the classics emphasizing the role of honey as a natural emulsifying agent in standardization of ‘Maadhutailika Basti’.
PMCID: PMC3215309  PMID: 22131675
Enema; Aasthaapana Basti; Basti dravya;  Emulsion stability; Emulsifying agent
6.  An experimental study to evaluate the pharmacokinetic aspect of Lekhana Basti (Emaciating/ Desiccating Medicated Enema) 
Ancient Science of Life  2011;31(2):38-43.
Basti therapy is used widely for treatment of various diseases in the field of Ayurveda with several proven clinical applications, however; the exact mechanism through which the benefits are obtained are yet to be ascertained in the light of the contemporary developments in clinical science. It is also not clear that when administered Basti the active principles in the formulation gain entry in to the systemic circulation or not. The present study was planned to acquire some preliminary data with regards to the absorption of phytochemical constituents of the formulations when administered in the form of Basti. Gallic acid was used as marker compound and it's absorption in systemic circulation was assessed using high performance liquid chromatography. Gallic acid present in Lekhana Basti (Emaciating/ Desiccating Medicated Enema) was found to get absorbed in to systemic circulation. Maximum concentration in serum was seen in the animal which received Lekhana Basti with Prakshepa Dravya ( Catalytic Adjuvant) in comparison to animal which received Lekhana Basti without Prakshepa Dravya. Area under curve in chromatogram was also more in animal which received Lekhana Basti with Prakshepa Dravya. From primary evidence created by HPLC study it can be said that, phytochemicals of the Basti formulation do get absorbed in systemic circulation.
PMCID: PMC3530265  PMID: 23284203
Lekhana Basti; Prakshepa Dravya; pharmacokinetic; Gallic acid; albino rats
7.  Antiosteoporotic activity of phytoestrogen-rich fraction separated from ethanol extract of aerial parts of Cissus quadrangularis in ovariectomized rats 
Indian Journal of Pharmacology  2012;44(3):345-350.
Cissus quadrangularis L. (C. quadrangularis L.) (Vitaceae) has been reported in Ayurveda for its antiosteoporotic activity. The study separated the phytoestrogen-rich fraction (IND-HE) from aerial parts of C. quadrangularis L. and evaluated its effect on osteoporosis caused by ovariectomy in rats.
Materials and Methods:
IND-HE was separated from the ethanol extract of C. quadrangularis. Ovariectomized female Wistar rats were divided into four groups (n = 6). Group 1: Control (distilled water), Group II: IND-HE (75 mg/kg p.o.), Group III: IND-HE (100 mg/kg p.o.) were treated once daily for 8 weeks and Group IV: standard estradiol group, received estrogen (1 mg/kg, s.c. bi-weekly). The effects on body weight were determined. DEXA (Dual energy-emission X-ray absorptimatory analysis) of whole body bone and femur was carried out. Blood was removed and analyzed for biochemical parameters. After sacrificing the animals, biomechanical study of right tibia and histopathology of pelvic bone was carried out.
IND-HE showed presence of phytoestrogen-rich fraction. IND-HE (75 and 100 mg/ kg) and estrogen treatment showed statistically significant increase in bone thickness, bone density and bone hardness. IND-HE (75 and 100 mg/kg) and estrogen treatment significantly increased serum estradiol. IND-HE (100 mg/kg) (P<0.05) and estrogen treatment increased serum vitamin D3 and serum calcium compared to control. Alkaline phosphatase was significantly reduced by IND-HE (100 mg/kg p.o.) and estrogen treatment. Histopathology and DEXA results indicated that IND-HE (75 and 100 mg/kg) prevented bone loss.
Discussion and Conclusion:
These findings confirm that phytoestrogen-rich fraction (IND- HE) possess good antiosteoporotic activity.
PMCID: PMC3371457  PMID: 22701244
Antiosteoporosis; estrogenic activity; phytoestrogen-rich fraction; ovariectomized rats; serum alcium
8.  Assessment of Bone Quality and Structure 
According to an NIH Consensus Conference in 2001, bone quality is related to various aspects of bone: its micro- and macroarchitecture, turnover, resorption and mineralisation. Radiological imaging techniques can be used to visualise and quantify bone micro- and macroarchitecture in vivo.
The parameters of bone macrostructure can be obtained using various methods: X-rays, DXA, QUS, QCT and MR imaging.
The parameters derived from traditional radiological investigations (such as hip axis length, neck width and neck shaft angle), like Singh indices, have been shown to be of limited usefulness in the diagnosis of osteoporosis and, indeed, have never been accepted as standard diagnostic tools.
Geometrical parameters (e.g. hip axis length, length and width of the neck of the femur) have also been obtained from DXA images of the hip; it has been shown that an increase, of two standard deviations, in hip axis length triples the risk of hip fracture. These measures have been used in numerous studies, but none has been shown, convincingly, to add substantial information to BMD in predicting status or fracture risk.
QUS provides quantitative parameters that are used to establish the properties of bone tissue. This method offers a series of advantages: smaller dimensions, simple and rapid measurements, no need for ionising radiations, as well as low cost compared with DXA and QCT. Consequently, the QUS seems to generate more information on bone fragility, to the extent that, at present, QUS systems are the ones most used in osteoporotic fracture risk prediction. Given the availability of various techniques for evaluating risk fracture, the T-score approach to fracture risk assessment seems to present some shortcomings linked to discrepancies between examined sites and techniques used. Ten-year fracture probability is the best method for determining the threshold for intervention.
QCT images, too, have been used to measure geometrical parameters; it was found that patients with osteoporotic fractures had a greater vertebral axial cross-section than fracture-free patients, and that treatment with parathormone increases the vertebral area. Initial studies have been performed using MRI data to generate geometrical parameters.
The microarchitectural parameters of trabecular bone structure have proved to be more useful than bone macroarchitecture measurements in evaluation of bone quality and in distinguishing between patients with and without osteoporotic fractures. Clinical studies have been performed using high-resolution techniques to study trabecular bone architecture; these techniques include multidetector CT, magnetic resonance and in vivo micro-CT. Indeed, using multidetector CT, bone structure measurements were shown to be better than BMD in differentiating between the two patient groups. However, the radiation dose needed to obtain sufficiently high quality images was found to be necessarily rather high, which is thus a potential limitation of this technique. HR-MR imaging, on the other hand, does not involve the use of radiation and is therefore more attractive for scientific studies. It has been used to study trabecular bone architecture in a number of studies, showing a good ability to discriminate between patients with and without osteoporotic fractures. The sites most frequently studied using HR-MR imaging are the distal radius and heel. The disadvantage of MR-based techniques is that the use of standard 1.5 Tesla systems is limited to peripheral parts, like the heel, distal tibia and distal radius, whereas higher magnetic fields (3 Tesla) would allow better visualisation of the trabecular bone structure and examination of more central parts of the skeleton, such as the proximal femur.
In vivo micro-CT is a recently developed imaging technique; initial studies on its ability to quantify the bone microarchitecture of the peripheral skeleton have given good results in terms of reproducibility and also capacity to detect age- and disease-related changes.
In conclusion, in vivo imaging of bone macro- and microarchitecture is possible, and a certain number of studies, geared at the optimisation and clinical application of these techniques, have already been conducted. The NIH in the USA are promoting and supporting the concept of bone quality, which in the future could lead to new diagnostic standards and techniques for analysing bone structure and will probably change the definition of osteoporosis.
PMCID: PMC3213808
9.  Inhibition of Bone Loss by Cissus quadrangularis in Mice: A Preliminary Report 
Journal of Osteoporosis  2012;2012:101206.
Women drastically loose bone during and after menopause leading to osteoporosis, a disease characterized by low bone mass increasing the risk of fractures with minor trauma. Existing therapies mainly reduce bone resorption, however, all existing drugs have severe side effects. Recently, the focus is to identify alternative medicines that can prevent and treat osteoporosis with minimal or no side effects. We used Cissus quadrangularis (CQ), a medicinal herb, to determine its effects on bone loss after ovariectomy in C57BL/6 mice. Two-month old mice were either sham operated or ovariectomized and fed CQ diet. After eleven weeks, mice were sacrificed and the long bones scanned using pQCT and μCT. In the distal femoral metaphysis, femoral diaphysis, and proximal tibia, control mice had decreased cancellous and cortical bone, while CQ-fed mice showed no significant differences in the trabecular number, thickness, and connectivity density, between Sham and OVX mice, except for cortical bone mineral content in the proximal tibia. There were no changes in the bone at the tibio-fibular junction between groups. We conclude that CQ effectively inhibited bone loss in the cancellous and cortical bones of femur and proximal tibia in these mice.
PMCID: PMC3388386  PMID: 22779034
10.  Evidence-based assessment of antiosteoporotic activity of petroleum-ether extract of Cissus quadrangularis Linn. on ovariectomy-induced osteoporosis 
Upsala Journal of Medical Sciences  2009;114(3):140-148.
The increasing incidence of postmenopausal osteoporosis and its related fractures have become global health issues in the recent days. Postmenopausal osteoporosis is the most frequent metabolic bone disease; it is characterized by a rapid loss of mineralized bone tissue. Hormone replacement therapy has proven efficacious in preventing bone loss but not desirable to many women due to its side-effects. Therefore we are in need to search the natural compounds for a treatment of postmenopausal symptoms in women with no toxic effects. In the present study, we have evaluated the effect of petroleum-ether extract of Cissus quadrangularis Linn. (CQ), a plant used in folk medicine, on an osteoporotic rat model developed by ovariectomy. In this experiment, healthy female Wistar rats were divided into four groups of six animals each. Group 1 was sham operated. All the remaining groups were ovariectomized. Group 2 was fed with an equivolume of saline and served as ovariectomized control (OVX). Groups 3 and 4 were orally treated with raloxifene (5.4 mg/kg) and petroleum-ether extract of CQ (500 mg/kg), respectively, for 3 months. The findings were assessed on the basis of animal weight, morphology of femur, and histochemical localization of alkaline phosphatase (ALP) (an osteoblastic marker) and tartrate-resistant acid phosphatase (TRAP) (an osteoclastic marker) in upper end of femur. The study revealed for the first time that the petroleum-ether extract of CQ reduced bone loss, as evidenced by the weight gain in femur, and also reduced the osteoclastic activity there by facilitating bone formation when compared to the OVX group. The osteoclastic activity was confirmed by TRAP staining, and the bone formation was assessed by ALP staining in the femur sections. The color intensity of TRAP and ALP enzymes from the images were evaluated by image analysis software developed locally. The effect of CQ was found to be effective on both enzymes, and it might be a potential candidate for prevention and treatment of postmenopausal osteoporosis. The biological activity of CQ on bone may be attributed to the phytogenic steroids present in it.
PMCID: PMC2852762  PMID: 19736603
Antiosteoporotic agents; ovariectomy; phytogenic steroid; postmenopausal osteoporosis
11.  The use of a Cissus quadrangularis/Irvingia gabonensis combination in the management of weight loss: a double-blind placebo-controlled study 
To evaluate the effects of two formulations, Cissus quadrangularis-only and a Cissus quadrangularis/Irvingia gabonensis combination, on weight loss in overweight and obese human subjects.
The study was a 10 week randomized, double-blind, placebo-controlled design involving 72 obese or overweight participants (45.8% male; 54.2% female; ages 21–44; mean age = 29.3). The participants were randomly divided into three equal (n = 24) groups: placebo, Cissus quadrangularis-only, and Cissus quadrangularis/Irvingia gabonensis combination. Capsules containing the placebo or active formulations were administered twice daily before meals; no major dietary changes nor exercises were suggested during the study. A total of six anthropomorphic and serological measurements (body weight, body fat, waist size; total plasma cholesterol, LDL cholesterol, fasting blood glucose level) were taken at baseline and at 4, 8 and 10 weeks.
Compared to the placebo group, the two active groups showed a statistically significant difference on all six variables by week 10. The magnitude of the differences was noticeable by week 4 and continued to increase over the trial period.
Although the Cissus quadrangularis-only group showed significant reductions on all variables compared to the placebo group, the Cissus quadrangularis/Irvingia gabonensis combination resulted in even larger reductions. This apparently synergistic formulation should prove helpful in the management of obesity and its related complications.
PMCID: PMC2330043  PMID: 18377661
12.  Management of tennis elbow by Agnikarma 
Tennis elbow is a painful condition and causes restricted movement of forearm which requires treatment for long period. Till date only symptomatic treatments are available like use of anti-inflammatory analgesic drugs, steroids injection, physiotherapy, exercise etc. But none of these provide satisfactory result. Long term use of anti-inflammatory, analgesic drugs and steroids injection is also not free from the adverse effects. Usually, ‘wait-and-see policy’ of treatment guideline is recommended in most of medical texts. According to Ayurveda, snayugata vata can be correlated with the condition of tennis elbow. Sushruta has advised Agnikarma for disorders of snayu (ligaments and tendons), asthi (bone), siddhi (joints) etc. Hence, in this study a case of tennis elbow (snayugata vata) was treated by Agnikarma, along with administration of powder of Ashwagandha and Navajivana Rasa orally, for a period of 03 weeks. This combination therapy provided considerable relief in pain and movement of the elbow joint.
PMCID: PMC3667434  PMID: 23741162
Agnikarma; Ashwagandha; Navajivana Rasa; snayugata vata; tennis elbow
13.  Petroleum Ether Extract of Cissus Quadrangularis (LINN) Stimulates the Growth of Fetal Bone during Intra Uterine Developmental Period: A Morphometric Analysis 
Clinics (Sao Paulo, Brazil)  2008;63(6):815-820.
The aim of the present study was to analyze the effect Cissus quadrangularis plant petroleum ether extract on the development of long bones during the intra-uterine developmental stage in rats.
Pregnant rats (n=12) were randomly assigned into either a control group (n=6) or a Cissus quadrangularis treatment (n=6) group. Pregnant rats in the Cissus quadrangularis group were treated with Cissus quadrangularis petroleum ether extract at a dose of 500 mg/kg body weight from gestation day 9 until delivery. The animals in the control group received an equal volume of saline. Newborn pups were collected from both groups for alizarin red S - alcian blue staining to differentiate ossified and unossified cartilage. The ossified cartilage (bone) was morphometrically analyzed using Scion image software.
Morphometric analysis revealed that the percentage of the total length of ossified cartilage (bone) in pups born to treated dams was significantly higher (P<0.001– 0.0001) than that of the control group.
The results of the present study suggest that maternal administration of Cissus quadrangularis petroleum ether extract during pregnancy can stimulate the development of fetal bone growth during the intra-uterine developmental period.
PMCID: PMC2664284  PMID: 19061006
Estrogens; Phytoestrogens; Ossification; Alizarin red-Alcian blue
14.  OA01.17. Efficacy of comprehensive ayurveda management of vertebral disc lesions by panchakarma therapies and herbomineral formulations 
Ancient Science of Life  2012;32(Suppl 1):S17.
Vertebral disc lesion i.e. spondylosis (Cervical / Thoracic /Lumbar) and PID affects nearly 80% of population. It is the most common cause of job related disability leading to missed work and second most common neurological ailment. As per the modern management is concerned, there is no established treatment for it, except surgical intervention, which cause complications and side effects. The present study has focus on the effectiveness of panchkarma combined with shamana therapies.
The study was conducted in 100 clinically diagnosed patients of vertebral disc lesions. These patients were randomly selected and divided in to two groups. I. Patients having cervical lesion were kept on Panchkarma (Udvartan and Valukasveda Snehan, Swedan, Vamankarm, Tiktksheer Basti, Greevabasti, Nasyakarma.) II. Patients having lumbar lesions ware kept on (Udvartan and Valukasveda Snehan, Swedan, Virechankarm, Tiktaksheer basti, Katibasti, Nasyakarma.)After that both groups were kept on oral drugs such as Tab. Arogyavardhini-250mg BID, and Tab. Panchtiktakgrithguggul-250mg BID. With local abhyanga and nadisveda for 1 month.
The patient was assessed after completion of treatment and follow up was done. The incidence of presenting features was worked out and the severities of the symptom were recorded statistically in each case and data were analysed. Both groups showed improvement in signs and symptoms significantly.
Vertebral disc lesions are progressive degenerative conditions in which vata at its own Ashaya (Asthi-Sandhi) gets provoked and as it belongs to Madhyam Rogamarga treatment becomes challenging. This attempt possibly finds solutions which can delay or even prevent surgical intervenation as sandhis are sites of Marma. It is an effective, simple, safe, treatment modality for vertebral disc lesion.
PMCID: PMC3800893
15.  Osteogenic potential of cissus qudrangularis assessed with osteopontin expression 
Fracture healing involves complex processes of cell and tissue proliferation and differentiation. Many factors are involved, including growth factors, inflammatory cytokines, antioxidants, bone breakdown (osteoclast) and bone building (osteoblast) cells, hormones, amino acids, and uncounted nutrients. We studied the osteogenic potential of Cissus quadrangularis (CQ), a plant that has been customarily used in the Indian subcontinent to hasten the process of healing in bone fractures.
Materials and Methods:
Total of 60 patients (age, 20-35 years) of mandible fracture was divided in two groups. Patients of group 1 were given capsules of CQ and fracture healing was assessed with osteopontin expression during treatment. Group 2 was control group.
Clinical and radiological analysis in our study was suggestive of better healing of fractures in group 1. All the samples of group 1 examined for osteopontin expression using western blot analysis and flow cytometry showed significant levels of expression of osteopontin protein and CD4+ T cells expressing osteopontin, respectively.
We conclude that CQ accelerates fracture healing and also causes early remodeling of fracture callus.
PMCID: PMC3800385  PMID: 24163553
Cissus quadrangularis; flow cytometry; osteopontin; western blot
16.  Strontium Ranelate: The Pathophysiological Rationale 
Skeletal metabolism and the replacement of damaged tissue with the same amount of intact bone depends on the correct balance between bone formation and bone resorption.
The existence of an imbalance between bone formation and resorption is a concept central to understanding of the pathophysiology of osteoporosis and the reduction of fracture risk.
With aging, the volume of bone that is formed during the bone remodelling process and after injury is less than the volume absorbed during the bone resorption phase; this results in bone loss and increased bone fragility. In addition to bone mineral density, many other properties of bone are determined by the balance between bone formation and bone resorption. A bone that is biomechanically more fragile is also a bone that consolidates more slowly after a fracture event. Although the fracture healing stages are the same even in the presence of osteoporosis, recent studies have shown a slowdown in the process of consolidation when osteoporosis is present. In particular, strategies to reduce fracture risk and facilitate the process of consolidation of the fracture may be a primary criterion for selection.
The ability to modulate anabolic and catabolic phenomena in the skeleton, both locally and systemically, opens up a new horizon for the reduction of fracture risk and the enhancement of bone healing, particularly when the bone is qualitatively and/or quantitatively compromised.
Clinical research has recently allowed the development of therapies, such as treatment with strontium ranelate, able to increase production of bone matrix by osteoblasts and to act positively on the distribution of the skeletal microarchitecture. Strontium ranelate is able to rebalance bone turnover in favour of the formation of more resistant and elastic bone, by stimulating osteoblasts and inhibiting the resorptive activity of osteoclasts, thereby ensuring rapid and lasting protection against the risk of fractures. In vitro studies have shown that the drug is able to promote replication of the first pre-osteoblasts and their differentiation into mature osteoblasts and osteocytes interacting with the receptor CaSR and through the increased synthesis of OPG. Thanks, again, to the participation of the CaSR receptor, but also by reducing the production of RANKL, strontium ranelate decreases the resorptive activity of osteoclasts. The anabolic action of strontium ranelate in terms of mineral apposition rate in both cortical and trabecular bone was demonstrated on bone biopsies analysed by three-dimensional micro-CT. The drug was shown to increase the number of trabeculae, the cortical thickness, and the total bone volume. The bone-forming activity of strontium ranelate was also demonstrated in comparative studies versus teriparatide and antiresorptive agents. In experimental studies the bone-forming effect of strontium ranelate leads to an increase in the bone callus volume and its maturation and, in turn, to an acceleration of the consolidation of the fracture and better implant osteointegration.
In conclusion, the mechanism of action of strontium ranelate, which inhibits bone resorption in favour of new bone formation, is able to counteract, in a physiological manner, the bone loss associated with advancing age. The net effect is an increase in bone mass, trabecular and cortical bone, which explains its anti-fracture efficacy. The drug’s ability to stimulate bone formation seems to unfold at the level of the callus allowing improved fracture healing and in the case of implants potential improvement of implant osteointegration.
PMCID: PMC3213810
17.  Clinical evaluation of cissus quadrangularis and moringa oleifera and osteoseal as osteogenic agents in mandibular fracture 
Ayurveda the ancient science of medicine describes various herb preparations that achieve the hastening of bone healing. Harjor showed clinical efficacy in the treatment of fractures.
The comparative evaluation of herbal agents as osteogenic agents in mandibular fractures.
Study design:
The patients were divided into four groups. Group 1: Osteoseal; Group 3: Harjor (Cissus quadrangularis); Group 2: Moringa (Moringa Oleifera); Group 4: Placebo.
Result and Conclusion:
Pain, Swelling, Tenderness, Mobility reduction is maximum in Osteoseal group and minimum in Placebo. There was an increase in the serum calcium and phosphorus level at different follow-ups in each groups but there was a decrease in the placebo group. Ca, Ca+, Phosphrous increase was maximum in the group 1.
PMCID: PMC3343389  PMID: 22639499
Osteogenic; prana; Asthisanghara
18.  Quantification of the Roles of Trabecular Microarchitecture and Trabecular Type in Determining the Elastic Modulus of Human Trabecular Bone 
The roles of microarchitecture and types of trabeculae in determining elastic modulus of trabecular bone have been studied in μCT images of 29 trabecular bone samples by comparing their Young’s moduli calculated by finite element analysis (FEA) with different trabecular type-specific reconstructions. The results suggest that trabecular plates play an essential role in determining elastic properties of trabecular bone.
Osteoporosis is an age-related disease characterized by low bone mass and architectural deterioration. Other than bone volume fraction (BV/TV), microarchitecture of bone is also believed to be important in governing mechanical properties of trabecular bone. We quantitatively examined the role of microarchitecture and relative contribution of trabecular types of individual trabecula in determining the elastic property of trabecular bone.
Materials and Methods
Twenty-nine human cadaveric trabecular bone samples were scanned at 21-μm resolution using a μCT system. Digital topological analysis (DTA) consisting of skeletonization and classification was combined with a trabecular type-specific reconstruction technique to extract the skeleton and identify topological type of trabeculae of the original trabecular bone image. Four different μCT-based finite element (FE) models were constructed for each specimen: (1) original full voxel; (2) skeletal voxel; (3) rod-reconstructed, preserving rod volume and plate skeleton; and (4) plate-reconstructed, preserving plate volume and rod skeleton. For each model, the elastic moduli were calculated under compression along each of three image-coordinate axis directions. Plate and rod tissue fractions directly measured from DTA-based topological classification were correlated with the elastic moduli computed from full voxel model.
The elastic moduli of skeleton models were significantly correlated with those of full voxel models along all three coordinate axes (r2 = 0.38~0.53). The rod-reconstructed model contained 21.3% of original bone mass and restored 1.5% of elastic moduli, whereas the plate-reconstructed model contained 90.3% of bone mass and restored 53.2% of elastic moduli. Plate tissue fraction showed a significantly positive correlation (r2 = 0.49) with elastic modulus by a power law, whereas rod tissue fraction showed a significantly negative correlation (r2 = 0.42).
These results quantitatively show that the microarchitecture alone affects elastic moduli of trabecular bone and trabecular plates make a far greater contribution than rods to the bone’s elastic behavior.
PMCID: PMC3225012  PMID: 16995816
osteoporosis; microarchitecture; trabecular type; digital topology; μCT imaging
19.  Development and Validation of a Simple Isocratic HPLC Method for Simultaneous Estimation of Phytosterols in Cissus quadrangularis 
Cissus quadrangularis L. is a promising remedy prescribed in the ancient Ayurvedic literature for bone fracture healing properties. As this activity has been extensively investigated and well established, a range of formulations containing C. quadrangularis has been marketed. This work reports the development and validation of a reliable RP-HPLC method for the analysis of phytosterols in the various extracts of the plant. The proposed method utilizes a Cosmosil C8 column (250 ΄ 4.6 mm) with a compatible Phenomenex C8 guard column with isocratic elution of acetonitrile and water (95:5 v/v) at 25°. An effluent flow rate of 2 ml/min and UV detection at 202 nm was used for the analysis of phytosterols. The described method was linear in the range of 1–500 μg/ml, with excellent correlation coefficients. The precision, robustness and ruggedness values were also within the prescribed limits (less than 2%). The recovery values were within the range, which indicates that the accuracy of the analysis was good and that the interference of the matrix with the recovery of phytosterols was low. The phytosterols were found to be stable in a stock solution for 48 h (% RSD was below 2%) and no interfering extra peaks were observed under controlled stress conditions. The proposed method is simple, specific, precise, accurate, and reproducible and thus can be used for routine analysis of C. quadrangularis phytosterols in quality control laboratories.
PMCID: PMC3178977  PMID: 21969748
Cissus quadrangularis; HPLC; phytosterols
20.  Clinical efficacy of Gokshura-Punarnava Basti in the management of microalbuminuria in diabetes mellitus 
Ayu  2012;33(4):537-541.
Microalbuminuria is the strong predictor of diabetic nephropathy, which is the main cause of morbidity and mortality in patients with diabetes mellitus (DM). Microalbuminuria is also characterized by increased prevalence of arterial hypertension, proliferative retinopathy, and peripheral neuropathy. The study was planned to evaluate the effect of Gokshura-Punarnava Basti in the management of microalbuminuria in DM (Madhumeha). Eligible diabetic patients with urine albumin excretion between 30 and 300 mg in 24 h were randomly divided into two groups. Asthapana Basti (decoction enema) of Gokshura and Punarnava Kwatha (decoction), Kalka (paste), Taila (medicated oil), Madhu (honey), and Saindhava (rock salt) for 6 consecutive days and Anuvasana (unctuous enema) of Gokshura-Punarnava Taila on 1st and 8th day by traditional Basti Putaka method was given in study group. Tablet Enalapril 5 mg, twice daily for 30 days was given to the patients in control group. The primary outcome measures were percentage change in the presenting complaints of diabetes, urine microalbumin, Blood Sugar Level (BSL), and Blood Pressure (BP). Enalapril showed 33.33% improvement, where as Gokshura-Punarnava Basti showed 79.59% improvement in the presenting complaints of diabetes, urine microalbumin, BSL and BP. Gokshura-Punarnava Basti has shown superior results in the management of microalbuminuria in DM as compared to control drug.
PMCID: PMC3665195  PMID: 23723672
Basti; diabetes mellitus; Gokshura; Madhumeha; microalbuminuria; Punarnava1
21.  The differential effects of bisphosphonates, SERMS (selective estrogen receptor modulators), and parathyroid hormone on bone remodeling in osteoporosis 
Osteoporosis is a skeletal metabolic disease characterized by a compromised bone fragility, leading to an increased risk of developing spontaneous and traumatic fractures. Osteoporosis is considered a multifactorial disease and fractures are the results of several different risk factors both extra- and intraskeletal. Thus bone fragility can be the end point of several different causes: a) failure to reach an optimal peak bone mass during growth; b) excessive bone resorption resulting in decreased bone mass and microarchitectural deterioration; c) inadequate formation upon an increased resorption during the process of bone remodeling. The pharmacological therapeutical options, available to date, are directed on prevention of fractures. The aim of this paper is to describe the activities and the mechanisms of action, as known at present, of the most used therapies for osteoporosis and their clinical implications. Improvement of knowledge in this field will allow us to further improve therapeutical choices and pharmacological interventions.
PMCID: PMC2684086  PMID: 18044075
Osteoporosis; estrogens; bisphosphonates; SERMS; teriparatide; mechanism of action; fracture
22.  Postmenopausal Osteoporosis: The Role of Immune System Cells 
In the last years, new evidences of the relationship between immune system and bone have been accumulated both in animal models and in humans affected by bone disease, such as rheumatoid arthritis, bone metastasis, periodontitis, and osteoporosis. Osteoporosis is characterized by low bone mass and microarchitectural deterioration of bone tissue with a subsequent increase in bone fragility and susceptibility to fractures. The combined effects of estrogen deprivation and raising of FSH production occurring in menopause cause a marked stimulation of bone resorption and a rapid bone loss which is central for the onset of postmenopausal osteoporosis. This review focuses on the role of immune system in postmenopausal osteoporosis and on therapeutic strategies targeting osteoimmunology pathways.
PMCID: PMC3677008  PMID: 23762093
23.  Effect of Shatapushpa Taila Matra Basti and Pathadi Kwatha on Poly Cystic Ovarian Disease 
Ayu  2012;33(2):243-246.
Poly Cystic Ovarian Disease (PCOD) is a complex disorder affecting 5-15% women in their reproductive age and related to ovarian dysfunction, characterized by menstrual irregularities, hyperandrogonism, obesity, and infertility. In Ayurveda, these symptoms are found under various conditions, caused by vitiated Vata and Kapha. Pathadi Kwatha and Shatapushpa Taila Matra Basti were studied in the current attempt to evaluate their comparative efficacy in cases of PCOD. Total 34 patients of PCOD were registered among which, 32 had completed the treatment of 2 months. They were randomly divided in to three groups. In group A Pathadi Kwatha (10 g, bid) and Shatapushpa Taila Matra Basti (60 ml for 7 days after cessation of menses for 2 consecutive cycles) were administered. In group B only Basti, whereas in group C capsules of roosted wheat flour were administered. Better results were obtained in group A especially in menstrual irregularities, achieving follicular growth and in weight reduction.
PMCID: PMC3611647  PMID: 23559797
Artavavaha Srotas; Matra Basti;  ovarian functions; Pathadi Kwatha;  PCOD; Shatapushpa Taila
24.  Effects of obesity on bone metabolism 
Obesity is traditionally viewed to be beneficial to bone health because of well-established positive effect of mechanical loading conferred by body weight on bone formation, despite being a risk factor for many other chronic health disorders. Although body mass has a positive effect on bone formation, whether the mass derived from an obesity condition or excessive fat accumulation is beneficial to bone remains controversial. The underline pathophysiological relationship between obesity and bone is complex and continues to be an active research area. Recent data from epidemiological and animal studies strongly support that fat accumulation is detrimental to bone mass. To our knowledge, obesity possibly affects bone metabolism through several mechanisms. Because both adipocytes and osteoblasts are derived from a common multipotential mesenchymal stem cell, obesity may increase adipocyte differentiation and fat accumulation while decrease osteoblast differentiation and bone formation. Obesity is associated with chronic inflammation. The increased circulating and tissue proinflammatory cytokines in obesity may promote osteoclast activity and bone resorption through modifying the receptor activator of NF-κB (RANK)/RANK ligand/osteoprotegerin pathway. Furthermore, the excessive secretion of leptin and/or decreased production of adiponectin by adipocytes in obesity may either directly affect bone formation or indirectly affect bone resorption through up-regulated proinflammatory cytokine production. Finally, high-fat intake may interfere with intestinal calcium absorption and therefore decrease calcium availability for bone formation. Unraveling the relationship between fat and bone metabolism at molecular level may help us to develop therapeutic agents to prevent or treat both obesity and osteoporosis.
Obesity, defined as having a body mass index ≥ 30 kg/m2, is a condition in which excessive body fat accumulates to a degree that adversely affects health [1]. The rates of obesity rates have doubled since 1980 [2] and as of 2007, 33% of men and 35% of women in the US are obese [3]. Obesity is positively associated to many chronic disorders such as hypertension, dyslipidemia, type 2 diabetes mellitus, coronary heart disease, and certain cancers [4-6]. It is estimated that the direct medical cost associated with obesity in the United States is ~$100 billion per year [7].
Bone mass and strength decrease during adulthood, especially in women after menopause [8]. These changes can culminate in osteoporosis, a disease characterized by low bone mass and microarchitectural deterioration resulting in increased bone fracture risk. It is estimated that there are about 10 million Americans over the age of 50 who have osteoporosis while another 34 million people are at risk of developing the disease [9]. In 2001, osteoporosis alone accounted for some $17 billion in direct annual healthcare expenditure.
Several lines of evidence suggest that obesity and bone metabolism are interrelated. First, both osteoblasts (bone forming cells) and adipocytes (energy storing cells) are derived from a common mesenchymal stem cell [10] and agents inhibiting adipogenesis stimulated osteoblast differentiation [11-13] and vice versa, those inhibiting osteoblastogenesis increased adipogenesis [14]. Second, decreased bone marrow osteoblastogenesis with aging is usually accompanied with increased marrow adipogenesis [15,16]. Third, chronic use of steroid hormone, such as glucocorticoid, results in obesity accompanied by rapid bone loss [17,18]. Fourth, both obesity and osteoporosis are associated with elevated oxidative stress and increased production of proinflammatory cytokines [19,20]. At present, the mechanisms for the effects of obesity on bone metabolism are not well defined and will be the focus of this review.
PMCID: PMC3141563  PMID: 21676245
bone; fat; obesity; osteoporosis; inflammation
25.  PA01.25. Clinical evaluation of chitrakadi churna and kshar basti in the management of amavata with special reference to rheumatoid arthritis 
Ancient Science of Life  2012;32(Suppl 1):S75.
The disease Amavata can be presented as very similar to Rheumatoid Arthritis. Rheumatoid Arthritis is chronic in nature and affects mostly middle aged people. For this disease there is no satisfactory medicine is available till date. But in Ayurveda there are many drugs described for Amavata which are cost effective and easily available with no side effects. The present clinical study evaluated the effect of Chitrakadi Churna and Kshar Basti in the management of Amavata.
30 clinically diagnosed patient of Amavata were registered and completed the trial. In this 15 patients (Group A) administered Chitrakadi Churna 4gm twice daily with lukewarm water after meal. Another 15 patients (Group B) administered Chitrakadi Churna 4gm twice daily with lukewarm water after meal and Kshar Basti as per Kalbasti krama i.e 16days followed by local Snehan Swedan.
Results of this trial were encouraging as there is improvement in each symptom of patient like pain, stiffness, swelling, and tenderness. Group A in which only Chitrakadi Churna was administered there was no significant improvement seen statistically. But Group B in which Chitrakadi Churna as well as Kshar Basti was administered highly significant improvement seen statistically. Details of the Statistical test and other important will be discussed at the time of paper presentation.
From present clinical trial it is concluded that this therapy is very useful for pain, swelling, tenderness and stiffness, which were chief complaint of the patient. Chronicity more than 3 years did not show marked improvement. And also this drug is supposed to be very good combination of Vedanashamaka, Shothaghna, Amapachaka Dravyas. No untoward effects were seen except mild loss of weight.
PMCID: PMC3800956

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