Biomedical subjects

Harutsugi Abukawa

Publications and source records attributed to Harutsugi Abukawa.

4 recordsLinked to original sources

PubMed2005

Hydrogel-beta-TCP scaffolds and stem cells for tissue engineering bone.

Trabecular bone is a material of choice for reconstruction after trauma and tumor resection and for correction of congenital defects. Autologous bone grafts are available in limited shapes and sizes; significant donor site morbidity is another major disadvantage to this approach. To overcome these limitations, we used a tissue engineering approach to create bone replacements in vitro, combining bone-marrow-derived differentiated mesenchymal stem cells (MSCs) suspended in hydrogels and 3-dimensionally printed (3DP) porous scaffolds made of beta-tricalcium-phosphate (beta-TCP). The scaffolds provided support for the formation of bone tissue in collagen I, fibrin, alginate, and pluronic F127 hydrogels during culturing in oscillating and rotating dynamic conditions. Histological evaluation including toluidine blue, alkaline phosphatase, and von Kossa staining was done at 1, 2, 4, and 6 weeks. Radiographic evaluation and high-resolution volumetric CT (VCT) scanning, expression of bone-specific genes and biomechanical compression testing were performed at 6 weeks. Both culture conditions resulted in similar bone tissue formation. Histologically collagen I and fibrin hydrogels specimens had superior bone tissue, although radiopacities were detected only in collagen I samples. VCT scan revealed density values in all but the Pluronic F127 samples, with Houndsfield unit values comparable to native bone in collagen I and fibrin glue samples. Expression of bone-specific genes was significantly higher in the collagen I samples. Pluronic F127 hydrogel did not support formation of bone tissue. All samples cultured in dynamic oscillating conditions had slightly higher mechanical strength than under rotating conditions. Bone tissue can be successfully formed in vitro using constructs comprised of collagen I hydrogel, MSCs, and porous beta-TCP scaffolds.

Alginates↗

PubMed2003

Formation of a mandibular condyle in vitro by tissue engineering.

PURPOSE: Mandibular reconstructive procedures often produce significant donor site morbidity. Recently, the use of minimally invasive techniques has been reported for mandibular reconstruction with decreased morbidity at the primary operative site. To date, these techniques have not addressed the graft donor site. We hypothesize that tissue-engineering techniques may be used to fabricate bone and thereby eliminate donor site morbidity. METHODS: Porcine mesenchymal stem cells (pMSCs) were isolated from the bone marrow of 3 Yucatan minipigs and grown in standard culture flasks. When they became near-confluent, cells were detached and replated with the addition of osteogenic supplements. A model of a porcine mandibular condyle was made and used to fabricate porous polymer scaffolds from biodegradable poly DL-lactic-co-glycolic acid (PLGA). Differentiated osteoblasts were transferred to the PLGA scaffold and cultured for 6 weeks in a rotational oxygen-permeable bioreactor system. The cultured constructs, consisting of scaffold and cells, were evaluated by gross, radiologic, and histologic examinations. RESULTS: The engineered constructs were white and hard and had a shape that closely resembled that of the model condyle. Plain radiographs demonstrated that the radiodensity of the construct was between that of the normal condyle and that of control scaffolds. Histologically, bone was observed on the entire surface of the PLGA scaffolds with an average thickness of 0.03 mm. Bone was not observed in the control scaffolds. CONCLUSION: In this pilot study, autologous tissue-engineered bone constructs were successfully made by combining biodegradable polymers and pMSCs.

Alkaline Phosphatase↗

PubMedDate not supplied

Osteoclastogenesis on tissue-engineered bone.

Bone remodeling plays an important role in bone function. To date, bone tissue-engineering research has focused primarily on bone formation from osteoblasts. This study demonstrates that osteoclastogenesis can occur on a mineralized polymer scaffold. Porcine bone marrow-derived mesenchymal stem cells (pMSCs) and hematopoietic cells were isolated from the bone marrow of Yucatan minipigs (n = 3) and cultured separately. pMSCs were differentiated into osteoblasts, seeded on porous poly(D,L-lactic-co-glycolic acid) foams, and cultured in a rotating oxygen-permeable bioreactor system. Once the cell-polymer constructs had started to mineralize, the hematopoietic cells were added and cocultured to include osteoclastogenesis. The cultured constructs were evaluated by histochemical and microscopic examination. Our results show that osteoblasts and osteoclasts were successfully differentiated from bone marrow on the scaffolds. This is the first demonstration of osteoclast formation on mineralized polymer surfaces.

Animals↗

PubMedDate not supplied

Tissue-engineered hybrid tooth and bone.

Tooth loss accompanied by alveolar bone resorption presents a significant clinical problem. We have investigated the utility of a tissue-engineering approach to provide corrective therapies for tooth-bone loss. Hybrid tooth-bone tissues were bioengineered as follows. Tooth implants were generated from pig third molar tooth bud cells seeded onto polyglycolide (PGA) and polyglycolide-colactide (PLGA) scaffolds, and grown for 4 weeks in the omenta of adult rat hosts. Bone implants were generated from osteoblasts induced from bone marrow progenitor cells obtained from the same pig, seeded onto PLGA fused wafer scaffolds, and grown for 10 days in a rotational oxygen-permeable bioreactor system. The tooth and bone implants were harvested, sutured together, reimplanted, and grown in the omenta for an additional 8 weeks. Histological and immunohistochemical analyses of the excised hybrid tooth-bone constructs revealed the presence of tooth tissues, including primary and reparative dentin and enamel in the tooth portion of hybrid tooth-bone implants, and osteocalcin and bone sialoprotein-positive bone in the bone portion of hybrid tooth-bone constructs. Collagen type III-positive connective tissue resembling periodontal ligament and tooth root structures were present at the interface of bioengineered tooth and bone tissues. These results demonstrate the utility of a hybrid tooth-bone tissue-engineering approach for the eventual clinical treatment of tooth loss accompanied by alveolar bone resorption.

Animals↗