Fractions of Shen-Sui-Tong-Zhi Formula Enhance Osteogenesis Via Activation of β-Catenin Signaling in Growth Plate Chondrocytes

Background: Shen-sui-tong-zhi formula (SSTZF) has been used to treat osteoporosis for decades and shows excellent clinical efficacy. This article aims to explore the optimal anti-osteoporotic ingredient and its precise mechanisms in mice models.Methods: In this study, we first screened the optimal anti-osteoporosis fraction of SSTZF extract in vivo, and then further explored the mechanism of its effects both in vivo and in vitro. Ten-week-old female C57BL/6J mice were administrated with each fraction of SSTZF. At 10 weeks after ovariectomy (OVX), femurs were collected for tissue analyses, including histology, micro-CT, biomechanical tests, and immunohistochemistry for ALP, FABP4, and β-catenin. Additionally, we also evaluated the mRNA expression level of ALP and FABP4 and the protein expression level of β-catenin after being treated with SSTZF extract in C3H10T1/2 cells. Moreover, we investigated the anti-osteoporosis effect of SSTZF extract on mice with β-catenin conditional knockout in growth plate chondrocytes (β-cateninGli1ER mice) through μCT, histology, and immunohistochemistry analyzes.Results: At 10 weeks after treatment, osteoporosis-like phenotype were significantly ameliorated in SSTZF n-butanol extract (SSTZF-NB) group mice, as indicated by increased trabecular bone area and ALP content, and decreased lipid droplet area and FABP4 content. No such improvements were observed after being treated with other extracts, demonstrating that SSTZF-NB is the optimal anti-osteoporosis fraction. Additionally, the elevated β-catenin was revealed in both OVX mice and C3H10T1/2 cells with SSTZF-NB administered. Furthermore, a significant osteoporosis-like phenotype was observed in β-cateninGli1ER mice as expected. However, SSTZF-NB failed to rescue the deterioration in β-cateninGli1ER mice, no significant re-upregulated ALP and downregulated FABP4 were observed after being treated with SSTZF-NB, demonstrating that SSTZF-NB prevents bone loss mainly via β-catenin signaling.Conclusion: SSTZF-NB enhances osteogenesis mainly via activation of β-catenin signaling in growth plate chondrocytes. SSTZF-NB is the optimal anti-osteoporosis fraction of SSTZF and it can be considered a salutary alternative therapeutic option for osteoporosis.

The Histomorphometry of Rabbit Bone Tissue with Experimental Osteoporosis after Implantation of Biphasic Calcium Phosphate Materials

The histomorphometry of the rabbit bone tissue from the lower jaw was done. Authors hypothesized that local enhancement with biphasic calcium phosphate ceramic materials in the femur trochanter major area increase the trabecular bone volume outside the implantation zone in vivo. Twenty-two California female rabbits were included in this study and were divided into four groups. Four healthy rabbits composed a control group (A group), while other eighteen underwent ovariectomy. Bone defects were created in femur trochanter major region. Sham surgery group (B group) consisted of four female rabbits with osteoporosis and bone defect, but no biomaterials were implanted. In C group (seven rabbits) created defects were filled with granules of biphasic calcium phosphate ceramic (hydroxyapatite (HAP) and tricalcium phosphate (TCP) 30/70); in D group (seven rabbits) defects were filled with the same granules (HAP/TCP 30/70) together with strontium (5% by mass). Twenty-two bone samples were taken from lower jaw premolar region. Trabecular bone area was measured using Image Pro Plus 7 program, where three equal fields (0.975 mm2) of view were at random chosen in all bone samples. Results have shown that the trabecular bone area in A group was 0.201 mm2 (0.176-0.233), which is statistically significantly higher (p <0.0001) than in B group 0.127 mm2 (0.118 – 0.149), C group 0.136 mm2 (0.108 – 0.166) and D group 0.135 mm2 (0.126 – 0.164), respectively. Statistically significant differences between B, C and D groups were not found (p > 0.05).

Effects of an antler velvet-based natural compound on osteoporosis in a rodent model

Context Velvet antlers (VA) are claimed to have the effects on osteoporosis in traditional Chinese medicine. Aims So as to scientifically confirm this claim, a VA-based compound (VAC, the mixture of upper part of VA, deer blood, and calcined oyster shell powder in a ratio of 4:1:1) was produced and administered to osteoporotic model rats, with osteoporosis being induced by retinoic acid via gavage. Methods In total, 48 rats were used and divided into six groups (8/group). Concentrations of alkaline phosphatase and osteocalcin in the rat serum were measured, and bone ash weight, concentrations of calcium and phosphorus in rat femur were measured. Bone biomechanical test was performed using a computer-controlled three-point bending tester. Ratio of trabecular bone area to tissue area in a given view field of the femoral tissue section was histologically examined and calculated. Bone histomorphometry was measured using micro-CT. Key results No significant difference was found between the VAC-treated groups and the positive control (alendronate sodium) on the basis of the following tested parameters: (1) levels of alkaline phosphatase and osteocalcin in rat serum; (2) maximum load value (N) of femur and maximum compression force of lumbar vertebra; (3) concentrations of calcium and phosphorus in femur; (4) ratio of trabecular bone area to tissue area; and (5) bone histomorphometry. Conclusions Effects of VAC used in the present study on osteoporosis in the model rats were comparable to the alendronate sodium (western medicine for treating osteoporosis) on the basis of our selected parameters. Implications This compound has the potential to be developed as an effective traditional Chinese medicine for clinic use to treat osteoporosis.

Effects of hydroxyapatite gypsum puger scaffold applied to rat alveolar bone sockets on osteoclasts, osteoblasts and the trabecular bone area

Background: Damage to bone tissue resulting from tooth extraction will cause alveolar bone resorption. Therefore, a material for preserving alveolar sockets capable of maintaining bone is required. Hydroxyapatite Gypsum Puger (HAGP) is a bio-ceramic material that can be used as an alternative material for alveolar socket preservation. The porous and rough surface of HAGP renders it a good medium for osteoblast cells to penetrate and attach themselves to. In general, bone mass is regulated through a remodeling process consisting of two phases, namely; bone formation by osteoblasts and bone resorption by osteoclasts. Purpose: This research aims to identify the effects of HAGP scaffold application on the number of osteoblasts and osteoclasts, as well as on the width of trabecular bone area in the alveolar sockets of rats. Methods: This research used Posttest Only Control Group Design. There were three research groups, namely: a group with 2.5% HAGP scaffold, a group with 5% HAGP scaffold and a group with 10% HAGP scaffold. The number of samples in each group was six. HAGP scaffold at concentrations of 2.5%, 5% and 10% was then mixed with PEG (Polyethylene Glycol). The Wistar rats were anesthetized intra-muscularly with 100 mg/ml of ketamine and 20 mg/ml of xylazine base at a ratio of 1:1 with a dose of 0.08-0.2 ml/kgBB. Extraction of the left mandibular incisor was performed before 0.1 ml preservation of HAGP scaffold + PEG material was introduced into the extraction sockets and suturing was performed. 7 days after preparation of the rat bone tissue, an Hematoxilin Eosin staining process was conducted in order that observation under a microscope could be performed. Results: There were significant differences in both the number of osteoclasts and osteoblasts between the 2.5% HAGP group, the 5% HAGP group and the 10% HAGP group (p = 0.000). Similarly, significant differences in the width of the trabecular bone area existed between the 5% HAGP group and the 10% HAGP group, as well as between the 2.5% HAGP group and the 10% HAGP group (p=0.000). In contrast, there was no significant difference in the width of the trabecular bone area between the 2.5% HAGP group and the 5% HAGP group. Conclusion: The application of HAGP scaffold can reduce osteoclasts, increase osteoblasts and extend the trabecular area in the alveolar bone sockets of rats.

Human Umbilical Cord Mesenchymal Stem-Cell Therapy to Increase the Density of Osteoporotic Mandibular Bone

Objective The aim of this study is to evaluate the feasibility of human umbilical cord mesenchymal stem-cell (hUCMSC) therapy in increasing osteoporotic mandibular bone density in a rat model by determining changes in alkaline phosphatase (ALP), osteocalcin, type 1 collagen, and trabecular bone area after treatment. Materials and Methods This research adopted an experimental posttest-only control group design. Thirty female Wistar rats were randomly divided into six groups, namely, a control group with rats postsham surgery (T1), osteoporotic model postovariectomy rats (T2), postovariectomy rats 4 weeks after gelatin injection (T3), postovariectomy rats 8 weeks after gelatin injection (T4), postovariectomy rats 4 weeks after hUCMSC injection (T5), and postovariectomy rats 8 weeks after hUCMSC injection (T6). The rats were all sacrificed for histological and immunohistochemical examinations of ALP, osteocalcin, type 1 collagen, and trabecular bone area. Results Increased expression of ALP, type 1 collagen, and osteocalcin, as well as increased trabecular bone area, was observed in the treatment groups compared with that in the osteoporotic groups. Conclusion hUCMSCs produce significant osteogenic effects and increase osteoporotic mandibular bone density in the animal model. Increases in bone density are demonstrated by the higher levels of ALP, osteocalcin, and type 1 collagen, as well as increases in the trabecular bone area.