J Korean Assoc Oral Maxillofac Surg.  2010 Apr;36(2):87-93. 10.5125/jkaoms.2010.36.2.87.

Maxillary sinus floor elevation using autogenous skin-derived mesenchymal stem cells in miniature pigs

• Byun JH
• Kang EJ
• Maeng GH
• Rho GJ
• Kang DH
• Lee JS
• Park BW

• KMID: 2189976
• DOI: http://doi.org/10.5125/jkaoms.2010.36.2.87

Abstract

INTRODUCTION
In our previous studies, we isolated porcine skin-derived mesenchymal stem cells (pSDMSCs) from the ears of adult miniature pigs and evaluated the pluripotency of these pSDMSCs based on expressions of transcription factors, such as Oct-4, Sox-2, and Nanog. Moreover, the characteristic of mesenchymal stem cells was revealed by the expression of various mesenchymal stem cell markers, including CD29, CD44, CD90, and vimentin. The aim of this study was to evaluate in vivo osteogenesis after maxillary sinus lift procedures with autogenous pSDMSCs and scaffold.
MATERIALS AND METHODS
The autogenous pSDMSCs were isolated from the 4 miniature pigs, and cultured to 3rd passage with same methods of our previous studies. After cell membranes were labeled using a PKH26, 1x10(7) cells/100 microliter of autogenous pSDMSCs were grafted into the maxillary sinus with a demineralized bone matrix (DBM) and fibrin glue scaffold. In the contralateral control side, only a scaffold was grafted, without SDMSCs. After two animals each were euthanized at 2 and 4 weeks after grafting, the in vivo osteogenesis was evaluated with histolomorphometric and osteocalcin immunohistochemical studies.
RESULTS
In vivo PKH26 expression was detected in all specimens at 2 and 4 weeks after grafting. Trabecular bone formation and osteocalcin expression were more pronounced around the grafted materials in the autogenous pSDMSCs-grafted group compared to the control group. Newly generated bone was observed growing from the periphery to the center of the grafted material.
CONCLUSION
The results of the present study suggest that autogenous skin-derived mesenchymal stem cells grafting with a DBM and fibrin glue scaffold can be a predictable method in the maxillary sinus floor elevation technique for implant surgery.

Keyword

Skin-derived mesenchymal stem cells; Tissue engineering; In vivo osteogenesis; Maxillary sinus floor elevation

MeSH Terms

Adult
Animals
Bone Matrix
Cell Membrane
Ear
Fibrin Tissue Adhesive
Floors and Floorcoverings
Humans
Maxillary Sinus
Mesenchymal Stromal Cells
Organic Chemicals
Osteocalcin
Osteogenesis
Swine
Tissue Engineering
Transcription Factors
Transplants
Vimentin
Fibrin Tissue Adhesive
Organic Chemicals
Osteocalcin
Transcription Factors
Vimentin

Figure

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  Fig. 1. Maxillary sinus floor elevation using autogenous porcine skin-derived mesenchymal stem cells. (pSDMSCs) A. The lateral window was formed via an extraoral approach in the anterior wall of the maxillary sinus. B. Augmented graft material was found under Schneider's membrane in the harvested specimen. (arrow)

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  Fig. 2. Cell labeling with PKH26. (A & B: 2 weeks after sinus grafting, x40 magnification; C & D: 4 weeks after grafting, x100 magnification) A. Numerous cells were observed in DAPI staining. B. PKH26 positive cells were plentifully detected in the same specimen. C, D. Abundant DAPI and PKH26 positive cells were also observed in the specimens of 4 weeks after sinus grafting.

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  Fig. 3. Histological features at 2 weeks after autogenous pSDMSCs grafting in the maxillary sinus floor. (A & C. x12 magnification; B & D, x40 magnification) A. Sinus opening and graft materials were observed in control specimen. B. High magnification of control specimen. Grafted DBM (arrow) was observed in the sinus elevated materials. However, new bone activity was barely detected in control group. C. Experimental group. The sinus grafted matrix was also detected in experimental group. Newly generated trabecular bones were clearly found in the peripheral portion of grafted material in the maxillary sinus. (arrows) D. High magnification of the experimental specimen, new bones were observed growing into the grafted materials.

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  Fig. 4. Histologic appearances of the in vivo generated new bones in the experimental specimens (A, × 40 magnification of 2 weeks specimen; B, x100 magnification of 2 weeks specimen; C & D, x40 magnification of 4 weeks specimens) A, B. New bone generation potential was detected in the specimens of 2 weeks after sinus grafting. C, D. More enhanced new bone generation and maturation were observed in the 4 weeks specimens compared with 2 weeks specimens. (Abbreviations: *, basal bone; arrow, borderline of basal bone and newly generated bone; arrow head, newly generated trabecular bone)

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  Fig. 5. Immunohistochemical staining of osteocalcin in the in vivo osteogenesis specimens. (A, control at 2 weeks; B, control at 4 weeks; C, experimental at 2 weeks; D, experimental at 4 weeks) (x40 magnification) A. Osteocalcin was almost negatively expressed in graft material except around DBM (arrow). B. Weak osteocalcin expression around newly generated bone (arrow) was found in the sinus graft material. C. Strong osteocalcin expressions were observed in the experimental specimens at both time points. D. Higher bone-morphogenic activity (arrow) was observed with enhanced osteocalcin expression in the peripheral portion of grafted material, especially in the immature fibroblast-like cells.

Reference

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