Conditions affecting cell seeding onto three-dimensional scaffolds for cellular-based biodegradable implants

Christian Weinand, Wei Xu Jian, Giuseppe M. Peretti, Lawrence J. Bonassar, Thomas J. Gill

Research output: Contribution to journalArticle

Abstract

Seeding cells efficiently and uniformly onto three-dimensional scaffolds is a key element for engineering tissues, particularly when only a low-number of cells is available for tissue repair and regeneration. The aim of this study was to evaluate three seeding techniques on two biocompatible scaffolds in vitro using chondrocytes as follows: (1) static; (2) modified centrifugal cell immobilization (CCI); and (3) dynamic oscillating motion. Five milliliters of media containing 5, 10, or 25 million articular, auricular, or costal chondrocytes were used to seed porous PLGA scaffolds and sections of devitalized cartilage. The dynamic oscillating technique resulted in up to 150% higher cellular load at 7 days than CCI seeding. Cell distribution was more homogeneous throughout the scaffold under dynamic conditions versus more sporadic and dispersed cell concentrations on the scaffolds when using either the static or the modified CCI technique. Cell load and distribution, when using a low numbers of chondrocytes at one and two million cells per milliliter, was comparable to that using the much higher number, especially under dynamic seeding conditions. The seeded scaffolds were used as implants to achieve cellular bonding between two devitalized meniscus discs. The constructs were implanted subcutaneously in nude mice for 12 weeks and analyzed histologically. Implants seeded with auricular chondrocytes showed qualitative more integration into native meniscus tissue than articular and costal cell implants. We conclude the dynamic oscillating seeding technique is an efficient technique for seeding low-cell numbers onto scaffolds resulting in consistent and uniform cell distribution throughout porous PLGA scaffolds.

Original languageEnglish
Pages (from-to)80-87
Number of pages8
JournalJournal of Biomedical Materials Research - Part B Applied Biomaterials
Volume91
Issue number1
DOIs
Publication statusPublished - Oct 2009

Fingerprint

Absorbable Implants
Scaffolds
Cell immobilization
Chondrocytes
Immobilization
Tissue
Cartilage
Cell Count
Joints
Scaffolds (biology)
Tissue engineering
Seed
Repair
Tissue Engineering
Nude Mice
Regeneration
Seeds

Keywords

  • Cell labeling
  • Chondrocyte
  • Dynamic-oscillating seeding
  • Implant
  • In vivo
  • Tissue engineering

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biomaterials

Cite this

Conditions affecting cell seeding onto three-dimensional scaffolds for cellular-based biodegradable implants. / Weinand, Christian; Jian, Wei Xu; Peretti, Giuseppe M.; Bonassar, Lawrence J.; Gill, Thomas J.

In: Journal of Biomedical Materials Research - Part B Applied Biomaterials, Vol. 91, No. 1, 10.2009, p. 80-87.

Research output: Contribution to journalArticle

@article{6d5a08bcfdd647d096113e7010f07531,
title = "Conditions affecting cell seeding onto three-dimensional scaffolds for cellular-based biodegradable implants",
abstract = "Seeding cells efficiently and uniformly onto three-dimensional scaffolds is a key element for engineering tissues, particularly when only a low-number of cells is available for tissue repair and regeneration. The aim of this study was to evaluate three seeding techniques on two biocompatible scaffolds in vitro using chondrocytes as follows: (1) static; (2) modified centrifugal cell immobilization (CCI); and (3) dynamic oscillating motion. Five milliliters of media containing 5, 10, or 25 million articular, auricular, or costal chondrocytes were used to seed porous PLGA scaffolds and sections of devitalized cartilage. The dynamic oscillating technique resulted in up to 150{\%} higher cellular load at 7 days than CCI seeding. Cell distribution was more homogeneous throughout the scaffold under dynamic conditions versus more sporadic and dispersed cell concentrations on the scaffolds when using either the static or the modified CCI technique. Cell load and distribution, when using a low numbers of chondrocytes at one and two million cells per milliliter, was comparable to that using the much higher number, especially under dynamic seeding conditions. The seeded scaffolds were used as implants to achieve cellular bonding between two devitalized meniscus discs. The constructs were implanted subcutaneously in nude mice for 12 weeks and analyzed histologically. Implants seeded with auricular chondrocytes showed qualitative more integration into native meniscus tissue than articular and costal cell implants. We conclude the dynamic oscillating seeding technique is an efficient technique for seeding low-cell numbers onto scaffolds resulting in consistent and uniform cell distribution throughout porous PLGA scaffolds.",
keywords = "Cell labeling, Chondrocyte, Dynamic-oscillating seeding, Implant, In vivo, Tissue engineering",
author = "Christian Weinand and Jian, {Wei Xu} and Peretti, {Giuseppe M.} and Bonassar, {Lawrence J.} and Gill, {Thomas J.}",
year = "2009",
month = "10",
doi = "10.1002/jbm.b.31376",
language = "English",
volume = "91",
pages = "80--87",
journal = "Journal of Biomedical Materials Research - Part A",
issn = "1549-3296",
publisher = "John Wiley and Sons Inc.",
number = "1",

}

TY - JOUR

T1 - Conditions affecting cell seeding onto three-dimensional scaffolds for cellular-based biodegradable implants

AU - Weinand, Christian

AU - Jian, Wei Xu

AU - Peretti, Giuseppe M.

AU - Bonassar, Lawrence J.

AU - Gill, Thomas J.

PY - 2009/10

Y1 - 2009/10

N2 - Seeding cells efficiently and uniformly onto three-dimensional scaffolds is a key element for engineering tissues, particularly when only a low-number of cells is available for tissue repair and regeneration. The aim of this study was to evaluate three seeding techniques on two biocompatible scaffolds in vitro using chondrocytes as follows: (1) static; (2) modified centrifugal cell immobilization (CCI); and (3) dynamic oscillating motion. Five milliliters of media containing 5, 10, or 25 million articular, auricular, or costal chondrocytes were used to seed porous PLGA scaffolds and sections of devitalized cartilage. The dynamic oscillating technique resulted in up to 150% higher cellular load at 7 days than CCI seeding. Cell distribution was more homogeneous throughout the scaffold under dynamic conditions versus more sporadic and dispersed cell concentrations on the scaffolds when using either the static or the modified CCI technique. Cell load and distribution, when using a low numbers of chondrocytes at one and two million cells per milliliter, was comparable to that using the much higher number, especially under dynamic seeding conditions. The seeded scaffolds were used as implants to achieve cellular bonding between two devitalized meniscus discs. The constructs were implanted subcutaneously in nude mice for 12 weeks and analyzed histologically. Implants seeded with auricular chondrocytes showed qualitative more integration into native meniscus tissue than articular and costal cell implants. We conclude the dynamic oscillating seeding technique is an efficient technique for seeding low-cell numbers onto scaffolds resulting in consistent and uniform cell distribution throughout porous PLGA scaffolds.

AB - Seeding cells efficiently and uniformly onto three-dimensional scaffolds is a key element for engineering tissues, particularly when only a low-number of cells is available for tissue repair and regeneration. The aim of this study was to evaluate three seeding techniques on two biocompatible scaffolds in vitro using chondrocytes as follows: (1) static; (2) modified centrifugal cell immobilization (CCI); and (3) dynamic oscillating motion. Five milliliters of media containing 5, 10, or 25 million articular, auricular, or costal chondrocytes were used to seed porous PLGA scaffolds and sections of devitalized cartilage. The dynamic oscillating technique resulted in up to 150% higher cellular load at 7 days than CCI seeding. Cell distribution was more homogeneous throughout the scaffold under dynamic conditions versus more sporadic and dispersed cell concentrations on the scaffolds when using either the static or the modified CCI technique. Cell load and distribution, when using a low numbers of chondrocytes at one and two million cells per milliliter, was comparable to that using the much higher number, especially under dynamic seeding conditions. The seeded scaffolds were used as implants to achieve cellular bonding between two devitalized meniscus discs. The constructs were implanted subcutaneously in nude mice for 12 weeks and analyzed histologically. Implants seeded with auricular chondrocytes showed qualitative more integration into native meniscus tissue than articular and costal cell implants. We conclude the dynamic oscillating seeding technique is an efficient technique for seeding low-cell numbers onto scaffolds resulting in consistent and uniform cell distribution throughout porous PLGA scaffolds.

KW - Cell labeling

KW - Chondrocyte

KW - Dynamic-oscillating seeding

KW - Implant

KW - In vivo

KW - Tissue engineering

UR - http://www.scopus.com/inward/record.url?scp=70049091367&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=70049091367&partnerID=8YFLogxK

U2 - 10.1002/jbm.b.31376

DO - 10.1002/jbm.b.31376

M3 - Article

C2 - 19388093

AN - SCOPUS:70049091367

VL - 91

SP - 80

EP - 87

JO - Journal of Biomedical Materials Research - Part A

JF - Journal of Biomedical Materials Research - Part A

SN - 1549-3296

IS - 1

ER -