Poly(L-Lactide)/Poly(∊-Caprolactone) and Collagen/β-Tricalcium Phosphate Scaffolds for the Treatment of Critical-Sized Rat Alveolar Defects: A Microtomographic,Molecular-Biological,and Histological Study
Restricted accessOtherFirst published online July, 2016
Poly(L-Lactide)/Poly(∊-Caprolactone) and Collagen/β-Tricalcium Phosphate Scaffolds for the Treatment of Critical-Sized Rat Alveolar Defects: A Microtomographic,Molecular-Biological,and Histological Study
To determine the efficacy of a newly developed scaffold (col/β-TCP) in a preclinical rat model as compared with the gold standard treatment (autograft) and control scaffolds (PLLA/PCL).
Design
Fifty-six Sprague-Dawley rats were randomized into four experimental groups, and critical-sized alveolar defects (7 × 4 × 3 mm) were created in each animal. Group A was the blank defect group, group B received autograft, group C received col/β-TCP scaffolds, and group D received PLLA/PCL blend scaffolds to fill the bone defects. New bone formation was assessed radiomorphometrically, histomorphometrically, and molecular-biologically at 1 and 4 months following surgery.
Results
Radiomorphometrically, the best new bone volume rate at 1 month (43.7%) and 4 months (45.4%) was observed in the autograft group, and the difference was significantly higher than in the other three groups (P ≤ .005, P ≤ .001, P ≤ .001 for 1 month and P = .004, P ≤ .001, P ≤ .001 for 4 months). Even though the new bone volume rate in the col/β-TCP group (21.5%) was higher than that of the PLLA/PCL group (18.2%), the difference was not significant (P = .08). Molecular-genetic analysis revealed significantly higher BSP and ALP gene expression levels in the autograft and col/β-TCP groups than in the blank defect group (P = .002 and P = .004).
Conclusion
The engineered tissue scaffolds described herein have great potential as an alternative treatment option when cost, donor region morbidity, and duration of hospitalization are considered.
AydinH.M., KorkusuzP., VargelI.Investigation of polymer/mesenchymal stem cell constructs for cranial defects: 6 months in vivo study. J Bioactive Compat Polym.2011; 26: 207–221.
2.
AydinH.M., YangY., KohlerT., HajA.E., MüllerR., PişkinE.Interaction of osteoblasts with macroporous scaffolds made of PLLA/PCL blends modified with collagen and hydroxyapatite. Adv Eng Mater.2009; 11: B83–B88.
BehniaH., KhojastehA., SoleimaniM., TehranchiA., KhoshzabanA., KeshelS.H., AtashiR.Secondary repair of alveolar clefts using human mesenchymal stem cells. Oral Surg Oral Med Oral Pathol Oral Radiol Endod.2009; 108: e1–e6.
5.
CalisM., DemirtasT.T., AtillaP., TatarI., ErsoyO., IrmakG., CelikH.H., CakarA.N., GumusdereliogluM., OzgurF.Estrogen as a novel agent for induction of adipose-derived mesenchymal stem cells for osteogenic differentiation: in vivo bone tissue-engineering study. Plast Reconstr Surg.2014; 133: 499e–510e.
6.
CalvertJ.W., WeissL.E., SundineM.J.New frontiers in bone tissue engineering. Clin Plast Surg.2003; 30: 641–648, x.
7.
ChurchmanS.M., KouroupisD., BoxallS.A., RoshdyT., TanH.B., McGonagleD., GiannoudisP.V., JonesE.A.Yield optimisation and molecular characterisation of uncultured CD271+ mesenchymal stem cells in the Reamer Irrigator Aspirator waste bag. Eur Cell Mater.2013; 26: 252–262.
8.
DasD.A multidisciplinary approach in treating a patient with unilateral cleft alveolus. Kathmandu Univ Med J (KUMJ).2013; 11: 171–174.
9.
DucyP., ZhangR., GeoffroyV., RidallA.L., KarsentyG.Osf2/Cbfa1: a transcriptional activator of osteoblast differentiation. Cell.1997; 89: 747–754.
LianJ.B., SteinG.S.Runx2/Cbfa1: a multifunctional regulator of bone formation. Curr Pharm Des.2003; 9: 2677–2685.
17.
LuM., RabieA.B.Quantitative assessment of early healing of intramembranous and endochondral autogenous bone grafts using micro-computed tomography and Q-win image analyzer. Int J Oral Maxillofac Surg.2004; 33: 369–376.
NewberryE.P., WillisD., LatifiT., BoudreauxJ.M., TowlerD.A.Fibroblast growth factor receptor signaling activates the human interstitial collagenase promoter via the bipartite Ets-AP1 element. Mol Endocrinol.1997; 11: 1129–1144.
20.
NguyenP.D., LinC.D., AlloriA.C., RicciJ.L., SaadehP.B., WarrenS.M.Establishment of a critical-sized alveolar defect in the rat: a model for human gingivoperiosteoplasty. Plast Reconstr Surg.2009; 123: 817–825.
21.
NguyenP.D., LinC.D., AlloriA.C., SchacharJ.S., RicciJ.L., SaadehP.B., WarrenS.M.Scaffold-based rhBMP-2 therapy in a rat alveolar defect model: implications for human gingivoperiosteoplasty. Plast Reconstr Surg.2009; 124: 1829–1839.
22.
OttoF., LubbertM., StockM.Upstream and downstream targets of RUNX proteins. J Cell Biochem.2003; 89: 9–18.
23.
Petrie AroninC.E., SadikK.W., LayA.L., RionD.B., TholpadyS.S., OgleR.C., BotchweyE.A.Comparative effects of scaffold pore size, pore volume, and total void volume on cranial bone healing patterns using microsphere-based scaffolds. J Biomed Mater Res A.2009; 89: 632–641.
24.
PfafflM.W., HorganG.W., DempfleL.Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res.2002; 30: e36.
25.
PrataC.A., LacerdaS.A., BrenteganiL.G.Autogenous bone graft associated with enamel matrix proteins in bone repair. Implant Dent.2007; 16: 413–420.
26.
QiH., AguiarD.J., WilliamsS.M., La PeanA., PanW., VerfaillieC.M.Identification of genes responsible for osteoblast differentiation from human mesodermal progenitor cells. Proc Natl Acad Sci U S A.2003; 100: 3305–3310.
27.
RachalaM.R., AileniK.R., DasariA.K., SinojiyaJ.Biomechanical considerations in mandibular incisor extraction cases. Int J Orthod Milwaukee.2015; 26: 47–51.
28.
RodriguesC.V., SerricellaP., LinharesA.B., GuerdesR.M., BorojevicR., RossiM.A., DuarteM.E., FarinaM.Characterization of a bovine collagen-hydroxyapatite composite scaffold for bone tissue engineering. Biomaterials.2003; 24: 4987–4997.
29.
RyooH.M., HoffmannH.M., BeumerT., FrenkelB., TowlerD.A., SteinG.S., SteinJ.L., van WijnenA.J., LianJ.B.Stage-specific expression of Dlx-5 during osteoblast differentiation: involvement in regulation of osteocalcin gene expression. Mol Endocrinol.1997; 11: 1681–1694.
30.
SarikayaB.A.H.Development of collagen/beta-TCP basedsynthetic bone grafts. J Tissue Eng Reg Med.2014; 8.
31.
SelvamuruganN., ChouW.Y., PearmanA.T., PulumatiM.R., PartridgeN.C.Parathyroid hormone regulates the rat collagenase-3 promoter in osteoblastic cells through the cooperative interaction of the activator protein-1 site and the runt domain binding sequence. J Biol Chem.1998; 273: 10647–10657.
32.
Shimizu-SasakiE., YamazakiM., FuruyamaS., SugiyaH., SodekJ., OgataY.Identification of a novel response element in the rat bone sialoprotein (BSP) gene promoter that mediates constitutive and fibroblast growth factor 2-induced expression of BSP. J Biol Chem.2001; 276: 5459–5466.
33.
ShuiC., SpelsbergT.C., RiggsB.L., KhoslaS.Changes in Runx2/Cbfa1 expression and activity during osteoblastic differentiation of human bone marrow stromal cells. J Bone Miner Res.2003; 18: 213–221.
34.
TodoM., ArahiraT.In vitro bone formation by mesenchymal stem cells with 3D collagen/beta-TCP composite scaffold. Conf Proc IEEE Eng Med Biol Soc.2013; 2013: 409–412.
35.
van HoutW.M., Mink van der MolenA.B., BreugemC.C., KooleR., Van CannE.M.Reconstruction of the alveolar cleft: can growth factor-aided tissue engineering replace autologous bone grafting? A literature review and systematic review of results obtained with bone morphogenetic protein-2. Clin Oral Invest.2011; 15: 297–303.
36.
WaiteP.D., WaiteD.E.Bone grafting for the alveolar cleft defect. Semin Orthod.1996; 2: 192–196.
37.
WangG.X., HuL., HuH.X., ZhangZ., LiuD.P.In vivo osteogenic activity of bone marrow stromal stem cells transfected with Ad-GFP-hBMP-2. Genet Mol Res.2014; 13: 4456–4465.
XiaoG., WangD., BensonM.D., KarsentyG., FranceschiR.T.Role of the alpha2-integrin in osteoblast-specific gene expression and activation of the Osf2 transcription factor. J Biol Chem.1998; 273: 32988–32994.
40.
YatabeM.S., OzawaT.O., JansonG., FacoR.A., GaribD.G.Are there bone dehiscences in maxillary canines orthodontically moved into the grafted alveolar cleft?Am J Orthod Dentofacial Orthop.2015; 147: 205–213.
41.
YilmazS., KilicA.R., KelesA., EfeogluE.Reconstruction of an alveolar cleft for orthodontic tooth movement. Am J Orthod Dentofacial Orthop.2000; 117: 156–163.
42.
ZhouH.Y., TakitaH., FujisawaR., MizunoM., KubokiY.Stimulation by bone sialoprotein of calcification in osteoblast-like MC3T3-E1 cells. Calcif Tissue Int.1995; 56: 403–407.
