Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Clinical Orthopaedics and Related Research®
Published in: Clinical Orthopaedics and Related Research® 8/2013

01-08-2013 | Symposium: Nanoscience in Musculoskeletal Medicine

Nano-ceramic Composite Scaffolds for Bioreactor-based Bone Engineering

Authors: Qing Lv, PhD, Meng Deng, PhD, Bret D. Ulery, PhD, Lakshmi S. Nair, M.Phil, PhD, Cato T. Laurencin, MD, PhD

Published in: Clinical Orthopaedics and Related Research® | Issue 8/2013

Login to get access

Abstract

Background

Composites of biodegradable polymers and bioactive ceramics are candidates for tissue-engineered scaffolds that closely match the properties of bone. We previously developed a porous, three-dimensional poly (D,L-lactide-co-glycolide) (PLAGA)/nanohydroxyapatite (n-HA) scaffold as a potential bone tissue engineering matrix suitable for high-aspect ratio vessel (HARV) bioreactor applications. However, the physical and cellular properties of this scaffold are unknown. The present study aims to evaluate the effect of n-HA in modulating PLAGA scaffold properties and human mesenchymal stem cell (HMSC) responses in a HARV bioreactor.

Questions/purposes

By comparing PLAGA/n-HA and PLAGA scaffolds, we asked whether incorporation of n-HA (1) accelerates scaffold degradation and compromises mechanical integrity; (2) promotes HMSC proliferation and differentiation; and (3) enhances HMSC mineralization when cultured in HARV bioreactors.

Methods

PLAGA/n-HA scaffolds (total number = 48) were loaded into HARV bioreactors for 6 weeks and monitored for mass, molecular weight, mechanical, and morphological changes. HMSCs were seeded on PLAGA/n-HA scaffolds (total number = 38) and cultured in HARV bioreactors for 28 days. Cell migration, proliferation, osteogenic differentiation, and mineralization were characterized at four selected time points. The same amount of PLAGA scaffolds were used as controls.

Results

The incorporation of n-HA did not alter the scaffold degradation pattern. PLAGA/n-HA scaffolds maintained their mechanical integrity throughout the 6 weeks in the dynamic culture environment. HMSCs seeded on PLAGA/n-HA scaffolds showed elevated proliferation, expression of osteogenic phenotypic markers, and mineral deposition as compared with cells seeded on PLAGA scaffolds. HMSCs migrated into the scaffold center with nearly uniform cell and extracellular matrix distribution in the scaffold interior.

Conclusions

The combination of PLAGA/n-HA scaffolds with HMSCs in HARV bioreactors may allow for the generation of engineered bone tissue.

Clinical Relevance

In cases of large bone voids (such as bone cancer), tissue-engineered constructs may provide alternatives to traditional bone grafts by culturing patients’ own MSCs with PLAGA/n-HA scaffolds in a HARV culture system.
Literature
1.
Attawia MA, Herbert KM, Uhrich KE, Langer R, Laurencin CT. Proliferation, morphology, and protein expression by osteoblasts cultured on poly(anhydride-co-imides). J Biomed Mater Res. 1999;48:322–327.PubMedCrossRef
2.
Berrey B, Lord C, Gebhardt M, Mankin H. Fractures of allografts. Frequency, treatment, and end-results. J Bone Joint Surg Am. 1990;72:825–833.PubMed
3.
Bianco P, Riminucci M, Gronthos S, Robey PG. Bone marrow stromal stem cells: nature, biology, and potential applications. Stem Cells. 2001;19:180–192.PubMedCrossRef
4.
Bil M, Ryszkowska J, Woźniak P, Kurzydłowski KJ, Lewandowska-Szumieł M. Optimization of the structure of polyurethanes for bone tissue engineering applications. Acta Biomaterialia. 2010;6:2501–2510.PubMedCrossRef
5.
Borden M, El-Amin SF, Attawia M, Laurencin CT. Structural and human cellular assessment of a novel microsphere-based tissue engineered scaffold for bone repair. Biomaterials. 2003;24:597–609.PubMedCrossRef
6.
Botchwey EA, Dupree MA, Pollack SR, Levine EM, Laurencin CT. Tissue engineered bone: Measurement of nutrient transport in three-dimensional matrices. J Biomed Mater Res A. 2003;67:357–367.PubMedCrossRef
7.
Botchwey EA, Pollack SR, El-Amin S, Levine EM, Tuan RS, Laurencin CT. Human osteoblast-like cells in three-dimensional culture with fluid flow. Biorheology. 2003;40:299–306.PubMed
8.
Botchwey EA, Pollack SR, Levine EM, Johnston ED, Laurencin CT. Quantitative analysis of three-dimensional fluid flow in rotating bioreactors for tissue engineering. J Biomed Mater Res A. 2004;69:205–215.PubMedCrossRef
9.
Botchwey EA, Pollack SR, Levine EM, Laurencin CT. Bone tissue engineering in a rotating bioreactor using a microcarrier matrix system. J Biomed Mater Res. 2001;55:242–253.PubMedCrossRef
10.
Brinker MR, O’Connor DP. The incidence of fractures and dislocations referred for orthopaedic services in a capitated population. J Bone Joint Surg Am. 2004;86:290–297.PubMed
11.
Bucaro M, Fertala J, Adams C, Steinbeck M, Ayyaswamy P, Mukundakrishnan K, Shapiro I, Risbud M. Bone cell survival in microgravity: evidence that modeled microgravity increases osteoblast sensitivity to apoptogens. Ann NY Acad Sci. 2004;1027:64–73.PubMedCrossRef
12.
Cai YZ, Zhang GR, Wang LL, Jiang YZ, Ouyang HW, Zou XH. Novel biodegradable three-dimensional macroporous scaffold using aligned electrospun nanofibrous yarns for bone tissue engineering. J Biomed Mater Res A. 2012;100:1187–1194.PubMed
13.
Cai Z, Zhang T, Di L, Xu DM, Xu DH, Yang DA. Morphological and histological analysis on the in vivo degradation of poly (propylene fumarate)/(calcium sulfate/β-tricalcium phosphate). Biomed Microdevices. 2011;13:623–631.PubMedCrossRef
14.
Chai YC, Roberts SJ, Schrooten J, Luyten FP. Probing the osteoinductive effect of calcium phosphate by using an in vitro biomimetic model. Tissue Eng Part A. 2010;17:1083–1097.CrossRef
15.
16.
Choi SW, Zhang Y, Thomopoulos S, Xia Y. In vitro mineralization by preosteoblasts in poly(DL-lactide-co-glycolide) inverse opal scaffolds reinforced with hydroxyapatite nanoparticles. Langmuir. 2010;26:12126–12131.PubMedCrossRef
17.
Ciapetti G, Granchi D, Baldini N. The combined use of mesenchymal stromal cells and scaffolds for bone repair. Curr Pharm Des. 2012;18:1796–1820.PubMedCrossRef
18.
Clarke S, Hoskins N, Jordan G, Marsh D. Healing of an ulnar defect using a proprietary TCP bone graft substitute, JAX‚ in association with autologous osteogenic cells and growth factors. Bone. 2007;40:939–947.PubMedCrossRef
19.
Dalby MJ, Gadegaard N, Tare R, Andar A, Riehle MO, Herzyk P, Wilkinson CDW, Oreffo ROC. The control of human mesenchymal cell differentiation using nanoscale symmetry and disorder. Nat Mater. 2007;6:997–1003.PubMedCrossRef
20.
Deng M, Kumbar SG, Lo KWH, Ulery BD, Laurencin CT. Novel polymer-ceramics for bone repair and regeneration. Recent Pat Biomed Eng. 2011;4:168–184.CrossRef
21.
Deng M, Kumbar SG, Nair LS, Weikel AL, Allcock HR, Laurencin CT. Biomimetic structures: biological implications of dipeptide-substituted polyphosphazene-polyester blend nanofiber matrices for load-bearing bone regeneration. Adv Funct Mater. 2011;21:2641–2651.CrossRef
22.
Deng M, Kumbar SG, Wan Y, Toti US, Allcock HR, Laurencin CT. Polyphosphazene polymers for tissue engineering: an analysis of material synthesis, characterization and applications. Soft Matter. 2010;6:3119–3132.CrossRef
23.
Deng M, Nair LS, Nukavarapu SP, Kumbar SG, Jiang T, Weikel AL, Krogman NR, Allcock HR, Laurencin CT. In situ porous structures: a unique polymer erosion mechanism in biodegradable dipeptide-based polyphosphazene and polyester blends producing matrices for regenerative engineering. Adv Funct Mater. 2010;20:2794–2806.CrossRef
24.
Dinopoulos H, Dimitriou R, Giannoudis PV. Bone graft substitutes: what are the options? Surgeon. 2012;10:230–239.PubMedCrossRef
25.
El-Amin S, Lu H, Khan Y, Burems J, Mitchell J, Tuan R, Laurencin C. Extracellular matrix production by human osteoblasts cultured on biodegradable polymers applicable for tissue engineering. Biomaterials. 2003;24:1213–1221.PubMedCrossRef
26.
Emani S, Mayer JE Jr, Emani SM. Gene regulation of extracellular matrix remodeling in human bone marrow stem cell-seeded tissue-engineered grafts. Tissue Eng A. 2011;17:2379–2388.CrossRef
27.
Erdemli O, Captug O, Bilgili H, Orhan D, Tezcaner A, Keskin D. In vitro and in vivo evaluation of the effects of demineralized bone matrix or calcium sulfate addition to polycaprolactone-bioglass composites. J Mater Sci Mat Med. 2010;21:295–308.CrossRef
28.
Gomes ME, Sikavitsas VI, Behravesh E, Reis RL, Mikos AG. Effect of flow perfusion on the osteogenic differentiation of bone marrow stromal cells cultured on starch-based three-dimensional scaffolds. J Biomed Mater Res A. 2003;67:87–95.PubMedCrossRef
29.
Grellier M, Bareille R, Bourget C, Amédée J. Responsiveness of human bone marrow stromal cells to shear stress. J Tissue Eng Regen Med. 2009;3:302–309.PubMedCrossRef
30.
Guelcher SA, Brown KV, Li B, Guda T, Lee BH, Wenke JC. Dual-purpose bone grafts improve healing and reduce infection. J Orthop Trauma. 2011;25:477–482.PubMedCrossRef
31.
Habibovic P, Van der Valk C, Van Blitterswijk C, De Groot K, Meijer G. Influence of octacalcium phosphate coating on osteoinductive properties of biomaterials. J Mater Sci Mater Med. 2004;15:373–380.PubMedCrossRef
32.
He W, Nieponice A, Soletti L, Hong Y, Gharaibeh B, Crisan M. Pericyte-based human tissue engineered vascular grafts. Biomaterials. 2010;31:8235–8244.PubMedCrossRef
33.
Ibim SEM, Uhrich KE, Attawia M, Shastri VR, El-Amin SF, Bronson R, Langer R, Laurencin CT. Preliminary in vivo report on the osteocompatibility of poly(anhydride-co-imides) evaluated in a tibial model. J Biomed Mater Res. 1998;43:374–379.PubMedCrossRef
34.
Ishaug SL, Crane GM, Miller MJ, Yasko AW, Yaszemski MJ, Mikos AG. Bone formation by three-dimensional stromal osteoblast culture in biodegradable polymer scaffolds. J Biomed Mater Res. 1997;36:17–28.PubMedCrossRef
35.
Ito H, Koefoed M, Tiyapatanaputi P, Gromov K, Goater JJ, Carmouche J, Zhang X, Rubery PT, Rabinowitz J, Samulski RJ, Nakamura T, Soballe K, O’Keefe RJ, Boyce BF, Schwarz EM. Remodeling of cortical bone allografts mediated by adherent rAAV-RANKL and VEGF gene therapy. Nat Med. 2005;11:291–297.PubMedCrossRef
36.
Jiang L, Li Y, Xiong C. Preparation and biological properties of a novel composite scaffold of nano-hydroxyapatite/chitosan/carboxymethyl cellulose for bone tissue engineering. J Biomed Sci. 2009;16:65.CrossRef
37.
Jiang T, Abdel-Fattah WI, Laurencin CT. In vitro evaluation of chitosan/poly(lactic acid-glycolic acid) sintered microsphere scaffolds for bone tissue engineering. Biomaterials. 2006;27:4894–4903.PubMedCrossRef
38.
Kondo N, Ogose A, Tokunaga K, Umezu H, Arai K, Kudo N, Hoshino M, Inoue H, Irie H, Kuroda K. Osteoinduction with highly purified beta-tricalcium phosphate in dog dorsal muscles and the proliferation of osteoclasts before heterotopic bone formation. Biomaterials. 2006;27:4419–4427.PubMedCrossRef
39.
Lao L, Wang Y, Zhu Y, Zhang Y, Gao C. Poly (lactide-co-glycolide)/hydroxyapatite nanofibrous scaffolds fabricated by electrospinning for bone tissue engineering. J Mater Sci Mater Med. 2011;22:1873–1884.PubMedCrossRef
40.
Laurencin CT, Ambrosio AMA, Borden MD, Cooper JA. Tissue engineering: orthopedic applications. Annu Rev Biomed Eng. 1999;1:19–46.PubMedCrossRef
41.
Lee JH, Rim NG, Jung HS, Shin H. Control of osteogenic differentiation and mineralization of human mesenchymal stem cells on composite nanofibers containing poly[lactic-co-(glycolic acid)] and hydroxyapatite. Macromol Biosci. 2010;10:173–182.PubMedCrossRef
42.
Lian JB, Stein GS. Concepts of osteoblast growth and differentiation: basis for modulation of bone cell development and tissue formation. Crit Rev Oral Biol Med. 1992;3:269–305.PubMed
43.
Lin L, Chow KL, Leng Y. Study of hydroxyapatite osteoinductivity with an osteogenic differentiation of mesenchymal stem cells. J Biomed Mater Res A. 2009;89:326–335.PubMed
44.
Lv Q, Nair L, Laurencin CT. Fabrication, characterization, and in vitro evaluation of poly(lactic acid glycolic acid)/nano-hydroxyapatite composite microsphere-based scaffolds for bone tissue engineering in rotating bioreactors. J Biomed Mater Res A. 2009;91:679–691.PubMed
45.
Marolt D, Campos IM, Bhumiratana S, Koren A, Petridis P, Zhang G, Spitalnik PF, Grayson WL, Vunjak-Novakovic G. Engineering bone tissue from human embryonic stem cells. Proc Natl Acad Sci. 2012;109:8705–8709.PubMedCrossRef
46.
Martinez E, Engel E, Planell J, Samitier J. Effects of artificial micro-and nano-structured surfaces on cell behaviour. Ann Anat. 2009;191:126–135.PubMedCrossRef
47.
Müller P, Bulnheim U, Diener A, Lüthen F, Teller M, Klinkenberg E-D, Neumann H-G, Nebe B, Liebold A, Steinhoff G, Rychly J. Calcium phosphate surfaces promote osteogenic differentiation of mesenchymal stem cells. J Cell Mol Med. 2008;12:281–291.PubMedCrossRef
48.
Ngiam M, Liao S, Patil AJ, Cheng Z, Chan CK, Ramakrishna S. The fabrication of nano-hydroxyapatite on PLGA and PLGA/collagen nanofibrous composite scaffolds and their effects in osteoblastic behavior for bone tissue engineering. Bone. 2009;45:4–16.PubMedCrossRef
49.
Pham QP, Sharma U, Mikos AG. Electrospun poly(ε-caprolactone) microfiber and multilayer nanofiber/microfiber scaffolds: characterization of scaffolds and measurement of cellular infiltration. Biomacromolecules. 2006;7:2796–2805.PubMedCrossRef
50.
Ripamonti U, Crooks J, Kirkbride A. Sintered porous hydroxyapatites with intrinsic osteoinductive activity: geometric induction of bone formation. South African J Sci. 1999;95:335–343.
51.
Saito E, Liao EE, Hu WW, Krebsbach PH, Hollister SJ. Effects of designed PLLA and 50: 50 PLGA scaffold architectures on bone formation in vivo. J Tissue Eng Regen Med. 2013;7:99–111.
52.
Salgado AJ, Coutinho OP, Reis RL. Bone tissue engineering: state of the art and future trends. Macromol Biosci. 2004;4:743–765.PubMedCrossRef
53.
Shao Z, Zhang X, Pi Y, Wang X, Jia Z, Zhu J, Dai L, Chen W, Yin L, Chen H. Polycaprolactone electrospun mesh conjugated with an MSC affinity peptide for MSC homing in vivo. Biomaterials. 2012;33:3375–3387.PubMedCrossRef
54.
Sikavitsas VI, Bancroft GN, Mikos AG. Formation of three-dimensional cell/polymer constructs for bone tissue engineering in a spinner flask and a rotating wall vessel bioreactor. J Biomed Mater Res. 2002;62:136–148.PubMedCrossRef
55.
Skardal A, Sarker SF, Crabbé A, Nickerson CA, Prestwich GD. The generation of 3-D tissue models based on hyaluronan hydrogel-coated microcarriers within a rotating wall vessel bioreactor. Biomaterials. 2010;31:8426–8435.PubMedCrossRef
56.
Thibault RA, Mikos AG, Kasper FK. Protein and mineral composition of osteogenic extracellular matrix constructs generated with a flow perfusion bioreactor. Biomacromolecules. 2011;12:4204–4212.PubMedCrossRef
57.
Vert M, Mauduit J, Li S. Biodegradation of PLA/GA polymers: increasing complexity. Biomaterials. 1994;15:1209–1213.PubMedCrossRef
58.
Xing Z, Xue Y, Dånmark S, Schander K, Ostvold S, Arvidson K, Hellem S, Finne-Wistrand A, Albertsson AC, Mustafa K. Effect of endothelial cells on bone regeneration using poly(L-lactide-co-1,5-dioxepan-2-one) scaffolds. J Biomed Mater Res A. 2011;96:349–357.PubMed
59.
Yang X, Wang D, Hao J, Gong M, Arlet V, Balian G, Shen FH, Li XJ. Enhancement of matrix production and cell proliferation in human annulus cells under bioreactor culture. Tissue Eng A. 2011;17:1595–1603.CrossRef
60.
Yu X, Botchwey EA, Levine EM, Pollack SR, Laurencin CT. Bioreactor-based bone tissue engineering: The influence of dynamic flow on osteoblast phenotypic expression and matrix mineralization. Proc Natl Acad Sci U S A. 2004;101:11203–11208.PubMedCrossRef
61.
Zandi M, Mirzadeh H, Mayer C, Urch H, Eslaminejad MB, Bagheri F, Mivehchi H. Biocompatibility evaluation of nano-rod hydroxyapatite/gelatin coated with nano-HAp as a novel scaffold using mesenchymal stem cells. J Biomed Mater Res A. 2010;92:1244–1255.PubMed
Metadata
Title
Nano-ceramic Composite Scaffolds for Bioreactor-based Bone Engineering
Authors
Qing Lv, PhD
Meng Deng, PhD
Bret D. Ulery, PhD
Lakshmi S. Nair, M.Phil, PhD
Cato T. Laurencin, MD, PhD
Publication date
01-08-2013
Publisher
Springer US
Published in
Clinical Orthopaedics and Related Research® / Issue 8/2013
Print ISSN: 0009-921X
Electronic ISSN: 1528-1132
DOI
https://doi.org/10.1007/s11999-013-2859-0

Other articles of this Issue 8/2013

Clinical Orthopaedics and Related Research® 8/2013 Go to the issue

Symposium: Nanoscience in Musculoskeletal Medicine

pHEMA-nHA Encapsulation and Delivery of Vancomycin and rhBMP-2 Enhances its Role as a Bone Graft Substitute

Symposium: Advanced Hip Arthroscopy

Novel CT-based Three-dimensional Software Improves the Characterization of Cam Morphology

Clinical Research

Does Early Functional Outcome Predict 1-year Mortality in Elderly Patients With Hip Fracture?

Obituary

Obituary: Kevin D. Harrington MD (1938–2013)

Editor's Spotlight/Take 5

Editor's Spotlight/Take 5: Nano-ceramic Composite Scaffolds for Bioreactor-based Bone Engineering

Clinical Research

Association Between Measures of Patella Height, Morphologic Features of the Trochlea, and Patellofemoral Joint Alignment: The MOST Study