PRELIMINARY STUDY ON STROMAL VASCULAR FRACTION PROMOTING ANGIOGENESIS AND TISSUE REGENERATION IN TISSUE ENGINEERING CHAMBER_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

LUFeng , CHANGQiang , ZHANWeiqing ,  LIXiaojian

Department of Plastic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou Guangdong, 510515, P. R. China;

Corresponding?author：

LIXiaojian, Email: lixj1999@hotmail.com

Keywords：

Adipose tissue engineering; Tissue engineering chamber; Stromal vascular fraction; Angiogenesis; Rabbit

DOI：

10.7507/1002-1892.20140142

Video：

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Objective To evaluate the mechanism of stromal vascular fraction (SVF) promoting angiogenesis and tissue regeneration in tissue engineering chamber. Methods Twenty-four 6-month-old New Zealand white rabbits, male or female, weighing 2.5-2.8 kg, were selected. Thoracic dorsal arteriovenous bundle combined with collagen type I scaffold was transplanted to dorsal side, and wrapped by cylindrical hollow silicone chamber; all animals were randomly divided into the experimental group (n=12) and the control group (n=12). SVF was isolated from the back fat pads of rabbits in experimental group and labelled with DiI at 2 weeks after operation. The 1 mL cell suspension (1×10⁶ cells/mL) and equal saline were injected into the chamber in experimental group and control group, respectively. The regenerative tissues were harvested for general observation and HE staining at 2 and 4 weeks after injection;and immunofluorescent staining was carried out in experimental group at 4 weeks. Results At 2 weeks after injection, the regenerative tissue was cylindrical; obvious vessel network and incompletely degradable collagen scaffold could be seen on the surface of the new tissue in 2 groups. The volume of new tissue was (0.87±0.11) mL in experimental group, and (0.72±0.08) mL in control group at 2 weeks, showing significant difference (t=2.701, P=0.011). At 4 weeks, little collagen scaffold could be seen on the surface in control group, but no collagen scaffold in experimental group; the volume of new tissue was (0.74±0.14) mL in experimental group, and (0.64±0.10) mL in control group, showing no significant difference (t=1.424, P=0.093). HE staining showed new mature vessels at 4 weeks, but no adipose tissue or fat lobulus formed in both groups; the capillary density was significantly higher in experimental group than in control group at 2 weeks (t=6.291, P=0.000) and at 4 weeks (t=5.445, P=0.000). The immunofluorescent staining found that SVF survived and located at the edge area after 4 weeks; the expressions of CD31 and DiI were positive in some endothelial cells. Conclusion SVF can promote the angiogenesis and tissue regeneration in tissue engineering chamber, but it can not differentiate into adipocyte spontaneously without adipogenic microenvironment.

Citation： LUFeng, CHANGQiang, ZHANWeiqing, LIXiaojian. PRELIMINARY STUDY ON STROMAL VASCULAR FRACTION PROMOTING ANGIOGENESIS AND TISSUE REGENERATION IN TISSUE ENGINEERING CHAMBER. Chinese Journal of Reparative and Reconstructive Surgery, 2014, 28(5): 644-648. doi: 10.7507/1002-1892.20140142 Copy

1.	Garvey PB, Buchel EW, Pockaj BA, et al. DIEP and pedicled TRAM flaps:a comparison of outcomes. Plast Reconstr Surg, 2006, 117(6):1720-1721.
2.	Tan S, Lim J, Yek J, et al. The deep inferior epigastric perforator and pedicled transverse rectus abdominis myocutaneous flap in breast reconstruction:a comparative study. Arch Plast Surg, 2013, 40(3):187-191.
3.	Patrick CW Jr, Chauvin PB, Hobley J, et al. Preadipocyte seeded PLGA scaffolds for adipose tissue engineering. Tissue Eng, 1999, 5(2):139-151.
4.	Cheung HK, Han TT, Marecak DM, et al. Composite hydrogel scaffolds incorporating decellularized adipose tissue for soft tissue engineering with adipose-derived stem cells. Biomaterials, 2014, 35(6):1914-1923.
5.	Hofer SO, Knight KM, Cooper-White JJ, et al. Increasing the volume of vascularized tissue formation in engineered constructs:an experimental study in rats. Plast Reconstr Surg, 2003, 111(3):1186-1192.
6.	Lilja HE, Morrison WA, Han XL, et al. An adipoinductive role of inflammation in adipose tissue engineering:key factors in the early development of engineered soft tissues. Stem Cells Dev, 2013, 22(10):1602-1613.
7.	Zuk PA, Zhu M, Mizuno H, et al. Multilineage cells from human adipose tissue:implications for cell-based therapies. Tissue Eng, 2001, 7(2):211-228.
8.	Zuk PA, Zhu M, Ashjian P, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell, 2002, 13(12):4279-4295.
9.	Bunnell BA, Flaat M, Gagliardi C, et al. Adipose-derived stem cells:isolation, expansion and differentiation. Methods, 2008, 45(2):115-120.
10.	Lu F, Mizuno H, Uysal CA, et al. Improved viability of random pattern skin flaps through the use of adipose-derived stem cells. Plast Reconstr Surg, 2008, 121(1):50-58.
11.	Gentile P, De Angelis B, Pasin M, et al. Adipose-derived stromal vascular fraction cells and platelet-rich plasma:basic and clinical evaluation for cell-based therapies in patients with scars on the face. J Craniofac Surg, 2014, 25(1):267-272.
12.	Gonzalez AM, Lobocki C, Kelly CP, et al. An alternative method for harvest and processing fat grafts:an in vitro study of cell viability and survival. Plast Reconstr Surg, 2007, 120(1):285-294.
13.	Sotorník R. Adipose tissue blood flow and metabolic syndrome. Cas Lek Cesk, 2010, 149(4):155-159.
14.	Kneser U, Kaufmann PM, Fiegel HC, et al. Long-term differentiated function of heterotopically transplanted hepatocytes on three-dimensional polymer matrices. J Biomed Mater Res, 1999, 47(4):494-503.
15.	Doldercr JH, Abberton KM, Thompson EW, et al. Spontaneous large volume adipose tissue generation from a vascularized pedicled fat flap inside a chamber space. Tissue Eng, 2007, 13(4):673-681.
16.	郝曉艷, 易成剛, 孟寶璽, 等. 組織工程室內帶血管蒂的脂肪瓣自發生長的研究. 中國美容醫學, 2010, 19(4):518-520.
17.	Matsuda K, Falkenberg KJ, Woods AA, et al. Adipose-derived stem cells promote angiogenesis and tissue formation for in vivo tissue engineering. Tissue Eng Part A, 2013, 19(11-12):1327-1335.
18.	Findlay MW, Dolderer JH, Trost N, et al. Tissue-engineered breast reconstruction:bridging the gap toward large-volume tissue engineering in humans. Plast Reconstr Surg, 2011, 128(6):1206-1215.
19.	Lokmic Z, Thomas JL, Morrison WA, et al. An endogenously deposited fibrin scaffold determines construct size in the surgically created arteriovenous loop chamber model of tissue engineering. J Vasc Surg, 2008, 48(4):974-985.

1. Garvey PB, Buchel EW, Pockaj BA, et al. DIEP and pedicled TRAM flaps:a comparison of outcomes. Plast Reconstr Surg, 2006, 117(6):1720-1721.
2. Tan S, Lim J, Yek J, et al. The deep inferior epigastric perforator and pedicled transverse rectus abdominis myocutaneous flap in breast reconstruction:a comparative study. Arch Plast Surg, 2013, 40(3):187-191.
3. Patrick CW Jr, Chauvin PB, Hobley J, et al. Preadipocyte seeded PLGA scaffolds for adipose tissue engineering. Tissue Eng, 1999, 5(2):139-151.
4. Cheung HK, Han TT, Marecak DM, et al. Composite hydrogel scaffolds incorporating decellularized adipose tissue for soft tissue engineering with adipose-derived stem cells. Biomaterials, 2014, 35(6):1914-1923.
5. Hofer SO, Knight KM, Cooper-White JJ, et al. Increasing the volume of vascularized tissue formation in engineered constructs:an experimental study in rats. Plast Reconstr Surg, 2003, 111(3):1186-1192.
6. Lilja HE, Morrison WA, Han XL, et al. An adipoinductive role of inflammation in adipose tissue engineering:key factors in the early development of engineered soft tissues. Stem Cells Dev, 2013, 22(10):1602-1613.
7. Zuk PA, Zhu M, Mizuno H, et al. Multilineage cells from human adipose tissue:implications for cell-based therapies. Tissue Eng, 2001, 7(2):211-228.
8. Zuk PA, Zhu M, Ashjian P, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell, 2002, 13(12):4279-4295.
9. Bunnell BA, Flaat M, Gagliardi C, et al. Adipose-derived stem cells:isolation, expansion and differentiation. Methods, 2008, 45(2):115-120.
10. Lu F, Mizuno H, Uysal CA, et al. Improved viability of random pattern skin flaps through the use of adipose-derived stem cells. Plast Reconstr Surg, 2008, 121(1):50-58.
11. Gentile P, De Angelis B, Pasin M, et al. Adipose-derived stromal vascular fraction cells and platelet-rich plasma:basic and clinical evaluation for cell-based therapies in patients with scars on the face. J Craniofac Surg, 2014, 25(1):267-272.
12. Gonzalez AM, Lobocki C, Kelly CP, et al. An alternative method for harvest and processing fat grafts:an in vitro study of cell viability and survival. Plast Reconstr Surg, 2007, 120(1):285-294.
13. Sotorník R. Adipose tissue blood flow and metabolic syndrome. Cas Lek Cesk, 2010, 149(4):155-159.
14. Kneser U, Kaufmann PM, Fiegel HC, et al. Long-term differentiated function of heterotopically transplanted hepatocytes on three-dimensional polymer matrices. J Biomed Mater Res, 1999, 47(4):494-503.
15. Doldercr JH, Abberton KM, Thompson EW, et al. Spontaneous large volume adipose tissue generation from a vascularized pedicled fat flap inside a chamber space. Tissue Eng, 2007, 13(4):673-681.
16. 郝曉艷, 易成剛, 孟寶璽, 等. 組織工程室內帶血管蒂的脂肪瓣自發生長的研究. 中國美容醫學, 2010, 19(4):518-520.
17. Matsuda K, Falkenberg KJ, Woods AA, et al. Adipose-derived stem cells promote angiogenesis and tissue formation for in vivo tissue engineering. Tissue Eng Part A, 2013, 19(11-12):1327-1335.
18. Findlay MW, Dolderer JH, Trost N, et al. Tissue-engineered breast reconstruction:bridging the gap toward large-volume tissue engineering in humans. Plast Reconstr Surg, 2011, 128(6):1206-1215.
19. Lokmic Z, Thomas JL, Morrison WA, et al. An endogenously deposited fibrin scaffold determines construct size in the surgically created arteriovenous loop chamber model of tissue engineering. J Vasc Surg, 2008, 48(4):974-985.

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Chinese Journal of Reparative and Reconstructive Surgery

PRELIMINARY STUDY ON STROMAL VASCULAR FRACTION PROMOTING ANGIOGENESIS AND TISSUE REGENERATION IN TISSUE ENGINEERING CHAMBER

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