Lithospermi radix extract-containing bilayer nanofiber scaffold for promoting wound healing in a rat model.

This study examined the in vitro characteristics and in vivo wound healing effect of novel Lithospermi radix (LR) extract-containing bilayer scaffolds in a rat model. LR extract, which has been used as a traditional herbal medicine for treating skin wounds, was added to a biocompatible gelatin solution. After glutaraldehyde vapor was used to modify the surface of chitosan scaffolds, various ratios of mammalian gelatin and fish collagen (GF100, GF91 and GF82) were electrospun onto the chitosan scaffolds to manufacture bilayer scaffolds. The porous chitosan scaffolds with a high swelling ratio showed efficient exudate absorption ability. GF91 gelatin nanofibers electrospun at a constant flow rate at 0.1 mL/h and a voltage of 20 kV displayed the optimal characteristics required for cell attachment and skin tissue regeneration. Moreover, the LR extract was successfully released slowly from the GF91 nanofibers. The investigation of the wound-healing activity of the chitosan/gelatin (CGF) bilayer scaffolds revealed that CGF91L provided the highest wound recovery rate in vivo in Sprague-Dawley (SD) rats. Based on its wound-healing effect and beneficial characteristics, the novel LR extract-containing CGF91 bilayer scaffold demonstrates potential as a material for treating skin wounds.

Enhanced Bone Tissue Regeneration by Porous Gelatin Composites Loaded with the Chinese Herbal Decoction Danggui Buxue Tang.

Danggui Buxue Tang (DBT) is a traditional Chinese herbal decoction containing Radix Astragali and Radix Angelicae sinensis. Pharmacological results indicate that DBT can stimulate bone cell proliferation and differentiation. The aim of the study was to investigate the efficacy of adding DBT to bone substitutes on bone regeneration following bone injury. DBT was incorporated into porous composites (GGT) made from genipin-crosslinked gelatin and ß-triclacium phosphates as bone substitutes (GGTDBT). The biological response of mouse calvarial bone to these composites was evaluated by in vivo imaging systems (IVIS), micro-computed tomography (micro-CT), and histology analysis. IVIS images revealed a stronger fluorescent signal in GGTDBT-treated defect than in GGT-treated defect at 8 weeks after implantation. Micro-CT analysis demonstrated that the level of repair from week 4 to 8 increased from 42.1% to 71.2% at the sites treated with GGTDBT, while that increased from 33.2% to 54.1% at GGT-treated sites. These findings suggest that the GGTDBT stimulates the innate regenerative capacity of bone, supporting their use in bone tissue regeneration.

Earthworm (Pheretima aspergillum) extract stimulates osteoblast activity and inhibits osteoclast differentiation.

BACKGROUND: The potential benefits of earthworm (Pheretima aspergillum) for healing have received considerable attention recently. Osteoblast and osteoclast activities are very important in bone remodeling, which is crucial to repair bone injuries. This study investigated the effects of earthworm extract on bone cell activities. METHODS: Osteoblast-like MG-63 cells and RAW 264.7 macrophage cells were used for identifying the cellular effects of different concentrations of earthworm extract on osteoblasts and osteoclasts, respectively. The optimal concentration of earthworm extract was determined by mitochondrial colorimetric assay, alkaline phosphatase activity, matrix calcium deposition, Western blotting and tartrate-resistant acid phosphatase activity. RESULTS: Earthworm extract had a dose-dependent effect on bone cell activities. The most effective concentration of earthworm extract was 3 mg/ml, significantly increasing osteoblast proliferation and differentiation, matrix calcium deposition and the expression levels of alkaline phosphatase, osteopontin and osteocalcin. Conversely, 3 mg/ml earthworm extract significantly reduced the tartrate-resistant acid phosphatase activity of osteoclasts without altering cell viability. CONCLUSIONS: Earthworm extract has beneficial effects on bone cell cultures, indicating that earthworm extract is a potential agent for use in bone regeneration.

Evaluating the bone tissue regeneration capability of the Chinese herbal decoction Danggui Buxue Tang from a molecular biology perspective.

Large bone defects are a considerable challenge to reconstructive surgeons. Numerous traditional Chinese herbal medicines have been used to repair and regenerate bone tissue. This study investigated the bone regeneration potential of Danggui Buxue Tang (DBT), a Chinese herbal decoction prepared from Radix Astragali (RA) and Radix Angelicae Sinensis (RAS), from a molecular biology perspective. The optimal ratio of RA and RAS used in DBT for osteoblast culture was obtained by colorimetric and alkaline phosphatase (ALP) activity assays. Moreover, the optimal concentration of DBT for bone cell culture was also determined by colorimetric, ALP activity, nodule formation, Western blotting, wound-healing, and tartrate-resistant acid phosphatase activity assays. Consequently, the most appropriate weight ratio of RA to RAS for the proliferation and differentiation of osteoblasts was 5:1. Moreover, the most effective concentration of DBT was 1,000 µg/mL, which significantly increased the number of osteoblasts, intracellular ALP levels, and nodule numbers, while inhibiting osteoclast activity. Additionally, 1,000 µg/mL of DBT was able to stimulate p-ERK and p-JNK signal pathway. Therefore, DBT is highly promising for use in accelerating fracture healing in the middle or late healing periods.

A novel porous gelatin composite containing naringin for bone repair.

As Gu-Sui-Bu (GSB) is a commonly used Chinese medical herb for therapeutic treatment of bone-related diseases, naringin is its main active component. This study elucidates how various concentrations of naringin solution affect the activities of bone cells, based on colorimetric, alkaline phosphatase activity, nodule formation, and tartrate-resistant acid phosphatase activity assays to determine the optimal concentration of naringin. GGT composite was obtained by combining genipin cross-linked gelatin and ß-tricalcium phosphate. GGTN composite was prepared by mixing GGT composite with the predetermined concentration of naringin. Porous GGT and GGTN composites were then made using a salt-leaching procedure. The potential of the composites in repairing bone defects was evaluated and compared in vivo by using the biological response of rabbit calvarial bone to these composites. Consequently, the most effective concentration of naringin was 10 mg/mL, which significantly enhanced the proliferation of osteoblasts, osteoclast activity, and nodule formation without affecting the alkaline phosphatase activity of osteoblasts and mitochondrial activity of mixed-bone cells. Radiographic analysis revealed greater new bone ingrowth in the GGTN composite than in the GGT composite at the same implantation time. Therefore, the GGTN composite is highly promising for use as a bone graft material.

Autologous bone marrow stromal cells loaded onto porous gelatin scaffolds containing Drynaria fortunei extract for bone repair.

GGT-GSB composite was prepared by mixing a biodegradable GGT composite containing genipin-crosslinked gelatin and ß-tricalcium phosphate with Gu-Sui-Bu extract (GSB) (Drynaria fortunei (Kunze) J. Sm.), a traditional Chinese medicine. Then, porous GGT and GGT-GSB scaffolds were fabricated using a salt-leaching method. The GGT and GGT-GSB scaffolds thus obtained had a macroporous structure and high porosity. Rabbit bone marrow stromal cells (BMSCs) were seeded onto GGT and GGT-GSB scaffolds. The biological response of rabbit calvarial bone to these scaffolds was considered to evaluate the potential of the scaffolds for use in bone tissue engineering. After 8 weeks of implantation, each scaffold induced new bone formation at a cranial bone defect, as was verified by X-ray microradiography. The BMSC-seeded GGT-GSB scaffolds induced more new bone formation than the BMSC-seeded GGT and acellular scaffolds. These observations suggest that an autologous BMSCs-seeded porous GGT-GSB scaffold can be adopted in bone engineering in vivo and has great potential for regenerating defective bone tissue.