Adipose-derived stem cells combined with inorganic bovine bone in calvarial bone healing in rats with type 2 diabetes.

BACKGROUND: Clinical studies have revealed that patients with type 2 diabetes mellitus (DM) have higher implant and bone grafting failure rates than the general population, likely owing to inferior bone healing. The authors sought to investigate whether adipose-derived stem cells (ASCs) combined with inorganic bovine bone improves bone repair in calvarial vertical critical-sized defects (CSDs) in rats with type 2 DM. METHODS: Bovine bone alone or seeded with 3 × 10(5), 3 × 10(6), or 3 × 10(7) ASCs/graft was randomly transplanted into calvarial CSDs in rats with DM induced by a high-fat diet with low-dose streptozotocin. Specimens were assayed using microcomputed tomography and histomorphometry at 4 and 8 weeks postimplantation. RESULTS: The histologic results showed an increase in new bone formation in the experimental groups compared with the control group. Both bone volume/total volume and trabecular thickness of newly formed bone within CSDs were the highest, and trabecular spacing was the lowest, in the 3 × 10(6) group at 8 weeks for the most favorable outcome. The results showed that the amount of new bone was greatest in the 3 × 10(6) group by 8 weeks. CONCLUSIONS: ASCs enhanced vertical bone regeneration in calvarial defects in rats with type 2 DM, when used in association with bovine bone scaffolds. The findings suggest that a combination of ASCs and bovine bone scaffolds could improve bone quantity in vertical bone defects.

In vitro culture and characterization of alveolar bone osteoblasts isolated from type 2 diabetics.

In order to understand the mechanisms of poor osseointegration following dental implants in type 2 diabetics, it is important to study the biological properties of alveolar bone osteoblasts isolated from these patients. We collected alveolar bone chips under aseptic conditions and cultured them in vitro using the tissue explants adherent method. The biological properties of these cells were characterized using the following methods: alkaline phosphatase (ALP) chemical staining for cell viability, Alizarin red staining for osteogenic characteristics, MTT test for cell proliferation, enzyme dynamics for ALP contents, radio-immunoassay for bone gla protein (BGP) concentration, and ELISA for the concentration of type I collagen (COL-I) in the supernatant. Furthermore, we detected the adhesion ability of two types of cells from titanium slices using non-specific immunofluorescence staining and cell count. The two cell forms showed no significant difference in morphology under the same culture conditions. However, the alveolar bone osteoblasts received from type 2 diabetic patients had slower growth, lower cell activity and calcium nodule formation than the normal ones. The concentration of ALP, BGP and COL-I was lower in the supernatant of alveolar bone osteoblasts received from type 2 diabetic patients than in that received from normal subjects (P < 0.05). The alveolar bone osteoblasts obtained from type 2 diabetic patients can be successfully cultured in vitro with the same morphology and biological characteristics as those from normal patients, but with slower growth and lower concentration of specific secretion and lower combining ability with titanium than normal ones.

Effects of local delivery of bFGF from PLGA microspheres on osseointegration around implants in diabetic rats.

OBJECTIVE: Diabetes mellitus may impair bone healing after dental implant placement. The objective of this study was to evaluate the effects of the local delivery of basic fibroblast growth factor (bFGF) from poly(lactide-co-glycolide) (PLGA) microspheres on osseointegration around titanium implants in diabetic rats. STUDY DESIGN: The bFGF-PLGA microspheres were prepared by the W/O/W double-emulsion solvent evaporation method. A total of 20 rats were used to create diabetic animal models by giving them a high-fat and high-sugar diet and a low-dose streptozotocin intraperitoneal injection. Titanium implants were planted into the tibias of the diabetic rats and into 10 normal rats. Microspheres were loaded on the surfaces of the implants in the bFGF intervention group before they were placed into the rats. After 4 or 8 weeks, the tibias containing the implants were removed and embedded with resin. Uncalcified tissue slices were prepared to compare osseointegration. RESULTS: At 4 weeks, the bone-implant contact rate in the diabetic control group was less than that in the control group and the bFGF intervention group (P < .05). At 8 weeks, the results among the 3 groups were similar to those at 4 weeks. CONCLUSIONS: The local delivery of bFGF from PLGA microspheres into areas around titanium implants may improve osseointegration in diabetic rats.

[Effect of local delivery of basic fibroblast growth factor 2 on osseointegration around implant in tibia of diabetic rats].

OBJECTIVE: To evaluate the effect of the local delivery of basic fibroblast growth factor 2 (bFGF-2) on the osseointegration around titanium implant of diabetic rats. METHODS: The bFGF-2-loaded poly (lactic-co-glycolic acid) microspheres were prepared by water/oil/water (W/O/W) double-emulsion solvent evaporation method. Thirty-five male SPF level Sprague Dawley rats, weighing 220-250 g and aged 9 weeks, were selected as experimental animals. Ten rats were fed with the routine diet as normal control group. The other 25 rats were made the diabetic animal model by giving high fat-sugar diet and a low dose streptozotocin (30 mg/kg) intravenously; 20 rats were made the diabetic animal model successfully. Then 20 rats were randomly divided into diabetic control group (n = 10) and bFGF-2 intervention group (n = 10). A hole was drilled in the right tibia bone of all rats, and the titanium implant treated by micro-arc oxidation surface was planted into the hole. Simultaneously, the previously prepared microspheres and blood were mixed and were loaded on the surface of the implant before it was implanted into the rats of the bFGF-2 intervention group. At 4 and 8 weeks, the tibia containing implants was harvested, embedded with resin and made undecalcified tissue slices to compare the osseointegration. RESULTS: At 4 weeks, the implants of the normal control group were surrounded by new lamellar bone with continuity; whereas the tissue around the implants of the diabetic control group contained little woven bone and some fibrous tissue; and obvious new formed bone with continuity was observed in bFGF-2 intervention group. At 8 weeks, the results of 3 groups were similar to those at 4 weeks. At 4 weeks, the percentage of bone-implant contact (BIC) in diabetic control group was significantly less than those in normal control group (P < 0.05) and in bFGF-2 intervention group (P < 0.05); the BIC in bFGF-2 intervention group was less than in normal control group, but showing no significant difference (P > 0.05). After 8 weeks, the BIC in normal control group and in bFGF-2 intervention group were significantly greater than that in diabetic control group (P < 0.05), but there was no significant difference between bFGF-2 intervention group and normal control group (P > 0.05). CONCLUSION: Local delivery of bFGF-2 around titanium implants may improve the osseointegration in diabetic rats.

Development of mesenchymal stem cell-implant complexes by cultured cells sheet enhances osseointegration in type 2 diabetic rat model.

This study investigated the hypothesis that a mesenchymal stem cells (MSC)-implant complex could be used in type 2 diabetic rats. Diabetes was modeled with type 2 diabetic rats induced by high fat diet with low dose streptozotocin (STZ) injected intraperitoneally. MSC sheets were harvested from culture flasks, wrapped around implants to construct the complexes, and then cultured in an osteogenic medium. The layered cell sheets integrated well with the implants and remained viable, with small mineralized nodules visible on the implant surfaces after culturing. The MSC-implant complexes were inserted into the right tibiae of the diabetic rats. Titanium implants served as controls. After four and eight weeks of healing, the tibiae were observed via MicroCT and harvested for histological examination. For the MSC-implant complexes, MicroCT analysis showed that bone volume ratio and trabecular thickness increased significantly (p<0.05), and trabecular separation decreased significantly (p<0.05) compared to the titanium implants in diabetic rats. Histological examination revealed a greater amount of new bone tissue forming around the MSC-implant complexes and a higher bone implant contact (BIC) rate than the titanium implants. These findings demonstrate that MSC-implant complexes possess osteogenic abilities and can be used in diabetic rats to improve the BIC rate. Thus, MSC-implant complexes provide a novel tissue engineering approach that promotes osseous healing and may potentially be useful in the treatment of diabetic patients.

In vitro culture and characterization of alveolar bone osteoblasts isolated from type 2 diabetics

In order to understand the mechanisms of poor osseointegration following dental implants in type 2 diabetics, it is important to study the biological properties of alveolar bone osteoblasts isolated from these patients. We collected alveolar bone chips under aseptic conditions and cultured them in vitro using the tissue explants adherent method. The biological properties of these cells were characterized using the following methods: alkaline phosphatase (ALP) chemical staining for cell viability, Alizarin red staining for osteogenic characteristics, MTT test for cell proliferation, enzyme dynamics for ALP contents, radio-immunoassay for bone gla protein (BGP) concentration, and ELISA for the concentration of type I collagen (COL-I) in the supernatant. Furthermore, we detected the adhesion ability of two types of cells from titanium slices using non-specific immunofluorescence staining and cell count. The two cell forms showed no significant difference in morphology under the same culture conditions. However, the alveolar bone osteoblasts received from type 2 diabetic patients had slower growth, lower cell activity and calcium nodule formation than the normal ones. The concentration of ALP, BGP and COL-I was lower in the supernatant of alveolar bone osteoblasts received from type 2 diabetic patients than in that received from normal subjects (P < 0.05). The alveolar bone osteoblasts obtained from type 2 diabetic patients can be successfully cultured in vitro with the same morphology and biological characteristics as those from normal patients, but with slower growth and lower concentration of specific secretion and lower combining ability with titanium than normal ones.