Secretory osteocalcin as a nondestructive osteogenic marker of tissue-engineered bone

Akahane, Manabu; Ueha, Tomoyuki; Dohi, Yoshiko; Shimizu, Takamasa; Tohma, Yasuaki; Kido, Akira; Kawate, Kenji; Imamura, Tomoaki; Tanaka, Yasuhito

doi:10.1007/s00776-011-0121-8

Cited by 7 publications

(17 citation statements)

References 23 publications

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“…7A, far right). For OCN, consistent with its known role as a late marker of osteogenesis [25], a more scattered distribution of positive staining was observed. This was most predominant within osteoid present in hPSC‐seeded scaffolds (Fig.…”

Section: Resultssupporting

confidence: 72%

An Abundant Perivascular Source of Stem Cells for Bone Tissue Engineering

James

Zara

Corselli

et al. 2012

Stem Cells Translational Medicine

119

152

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Adipose tissue is an ideal mesenchymal stem cell (MSC) source, as it is dispensable and accessible with minimal morbidity. However, the stromal vascular fraction (SVF) of adipose tissue is a heterogeneous cell population, which has disadvantages for tissue regeneration. In the present study, we prospectively purified human perivascular stem cells (PSCs) from n ‫؍‬ 60 samples of human lipoaspirate and documented their frequency, viability, and variation with patient demographics. PSCs are a fluorescence-activated cell sorting-sorted population composed of pericytes (CD45−, CD146؉, CD34−) and adventitial cells (CD45−, CD146−, CD34؉), each of which we have previously reported to have properties of MSCs. Here, we found that PSCs make up, on average, 43.2% of SVF from human lipoaspirate (19.5% pericytes and 23.8% adventitial cells). These numbers were minimally changed by age, gender, or body mass index of the patient or by length of refrigerated storage time between liposuction and processing. In a previous publication, we observed that human PSCs (hPSCs) formed significantly more bone in vivo in comparison with unsorted human SVF (hSVF) in an intramuscular implantation model. We now extend this finding to a bone injury model, observing that purified hPSCs led to significantly greater healing of mouse critical-size calvarial defects than hSVF (60.9% healing as opposed to 15.4% healing at 2 weeks postoperative by microcomputed tomography analysis). These studies suggest that adipose-derived hPSCs are a new cell source for future efforts in skeletal regenerative medicine. Moreover, hPSCs are a stem cell-based therapeutic that is readily approvable by the U.S. Food and Drug Administration, with potentially increased safety, purity, identity, potency, and efficacy. STEM CELLS TRANSLATIONAL MEDICINE 2012;1:673-684

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Section: Resultssupporting

confidence: 72%

An Abundant Perivascular Source of Stem Cells for Bone Tissue Engineering

James

Zara

Corselli

et al. 2012

Stem Cells Translational Medicine

119

152

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“…30 Combining BMSCs with scaffolds, such as β-TCP or HA, can promote the formation of new bone tissue after subcutaneous transplantation. 2,5,16-18 We also previously reported the osteogenic properties of HA constructs combined with BMSCs 1,5 and HA scaffolds wrapped with BMSC cell sheets. 22 Based on these results, we believe that cell sheet transplantation can be a useful method for making tissue-engineered bone for clinical applications.…”

Section: Discussionmentioning

confidence: 84%

“…Mesenchymal stem cells (MSCs) are adult stem cells which can be isolated from tissues such as bone marrow, 1,2 adipose, 3 and skeletal muscle. 4 Because MSCs are multipotent and can differentiate into osteoblasts, 2,5 chondrocytes, 6 and neurons, 7,8 they have been used in regenerative medicine to treat various diseases and tissue injuries, 9,10 including, tissue engineering using an in vitro cell culture.…”

Section: Introductionmentioning

confidence: 99%

Culturing bone marrow cells with dexamethasone and ascorbic acid improves osteogenic cell sheet structure

Akahane

Shimizu

Kira

et al. 2016

Bone & Joint Research

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ObjectivesTo assess the structure and extracellular matrix molecule expression of osteogenic cell sheets created via culture in medium with both dexamethasone (Dex) and ascorbic acid phosphate (AscP) compared either Dex or AscP alone.MethodsOsteogenic cell sheets were prepared by culturing rat bone marrow stromal cells in a minimal essential medium (MEM), MEM with AscP, MEM with Dex, and MEM with Dex and AscP (Dex/AscP). The cell number and messenger (m)RNA expression were assessed in vitro, and the appearance of the cell sheets was observed after mechanical retrieval using a scraper. β-tricalcium phosphate (β-TCP) was then wrapped with the cell sheets from the four different groups and subcutaneously implanted into rats.ResultsAfter mechanical retrieval, the osteogenic cell sheets from the MEM, MEM with AscP, and MEM with Dex groups appeared to be fragmented or incomplete structures. The cell sheets cultured with Dex/AscP remained intact after mechanical retrieval, without any identifiable tears. Culture with Dex/AscP increased the mRNA and protein expression of extracellular matrix proteins and cell number compared with those of the other three groups. More bridging bone formation was observed after transplantation of the β-TCP scaffold wrapped with cell sheets cultured with Dex/AscP, than in the other groups.ConclusionsThese results suggest that culture with Dex/AscP improves the mechanical integrity of the osteogenic cell sheets, allowing retrieval of the confluent cells in a single cell sheet structure. This method may be beneficial when applied in cases of difficult tissue reconstruction, such as nonunion, bone defects, and osteonecrosis.Cite this article: M. Akahane, T. Shimizu, T. Kira, T. Onishi, Y. Uchihara, T. Imamura, Y. Tanaka. Culturing bone marrow cells with dexamethasone and ascorbic acid improves osteogenic cell sheet structure. Bone Joint Res 2016;5:569–576. DOI: 10.1302/2046-3758.511.BJR-2016-0013.R1.

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“…Briefly, the β -TCP disks were wrapped with the OMCS and then autologously implanted atsubcutaneous site on the back of the rabbits [15, 21]. The β -TCP disks alone were also implanted as a control.…”

Section: Methodsmentioning

confidence: 99%

Osteogenic Matrix Cell Sheet Transplantation Enhances Early Tendon Graft to Bone Tunnel Healing in Rabbits

Inagaki

Uematsu

Akahane

et al. 2013

BioMed Research International

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The objective of this study was to determine whether osteogenic matrix cell sheets (OMCS) could induce bone formation around grafted tendons, thereby enhancing early stage tendon to bone tunnel healing in skeletally mature male Japanese white rabbits. First, the osteogenic potential of rabbit OMCS was evaluated. Then, the OMCS were transplanted into the interface between the grafted tendon and the bone tunnel created at the tibia. Histological assessments and biomechanical tensile testing were performed after 3 weeks. The rabbit OMCS showed high alkaline phosphatase (ALP) activity, positive staining of ALP, and osteogenic potential when transplanted subcutaneously with beta tricalcium phosphate disks. Newly formed bony walls and positive collagen type I staining were seen around the grafted tendon with OMCS transplantation, whereas such bony walls were thinner or less frequent without OMCS transplantation. Micro-computed tomography images showed significantly higher bone volume in the OMCS transplantation group. The pullout strength was significantly higher with OMCS (0.74 ± 0.23 N/mm2) than without OMCS (0.58 ± 0.15 N/mm2). These results show that OMCS enhance early tendon to bone tunnel healing. This method can be applied to cases requiring early tendon to bone tunnel healing after ligament reconstruction surgery.

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Secretory osteocalcin as a nondestructive osteogenic marker of tissue-engineered bone

Cited by 7 publications

References 23 publications

An Abundant Perivascular Source of Stem Cells for Bone Tissue Engineering

An Abundant Perivascular Source of Stem Cells for Bone Tissue Engineering

Culturing bone marrow cells with dexamethasone and ascorbic acid improves osteogenic cell sheet structure

Osteogenic Matrix Cell Sheet Transplantation Enhances Early Tendon Graft to Bone Tunnel Healing in Rabbits

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