Quantification of DNA in Biologic Scaffold Materials

Gilbert, Thomas W.; Freund, John M.; Badylak, Stephen F.

doi:10.1016/j.jss.2008.02.013

Cited by 438 publications

(382 citation statements)

References 42 publications

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“…The DNA content was used as an indicator of the presence of residual cells in decellularized tissue [28]. The DNA from either native or decellularized tissue samples of known masses (approximately 20 mg) was isolated, purified and rehydrated per triplicate using the Wizard® Genomic DNA Purification Kit (Promega) according to the recommended protocol.…”

Section: Deoxyribonucleic Acid (Dna) Isolation and Quantificationmentioning

confidence: 99%

Decellularization of pericardial tissue and its impact on tensile viscoelasticity and glycosaminoglycan content

et al. 2011

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Keywords:Tissue decellularization Pericardium Glycosaminoglycans Tensile properties Detergents Bovine pericardium is a collagenous tissue commonly used as a natural biomaterial in the fabrication of cardiovascular devices. For tissue engineering purposes, this xenogeneic biomaterial must be decellularized to remove cellular antigens. With this in mind, three decellularization protocols were compared in terms of their effectiveness to extract cellular materials, their effect on glycosaminoglycan (GAG) content and, finally, their effect on tensile biomechanical behavior. The tissue decellularization was achieved by treatment with t-octyl phenoxy polyethoxy ethanol (Triton X-100), tridecyl polyethoxy ethanol (ATE) and alkaline treatment and subsequent treatment with nucleases (DNase/RNase). The quantified residual DNA content (3.0 ± 0.4%, 4.4 ± 0.6% and 5.6 ± 0.7% for Triton X-100, ATE and alkaline treatment, respectively) and the absence of nuclear structures (hematoxylin and eosin staining) were indicators of effective cell removal. In the same way, it was found that the native tissue GAG content decreased to 61.6 ± 0.6%, 62.7 ± 1.1% and 88.6 ±0.2% for Triton X-100, ATE and alkaline treatment, respectively. In addition, an alteration in the tissue stress relaxation characteristics was observed after alkaline treatment. We can conclude that the three decellularization agents preserved the collagen structural network, anisotropy and the tensile modulus, tensile strength and maximum strain at failure of native tissue.

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Section: Deoxyribonucleic Acid (Dna) Isolation and Quantificationmentioning

confidence: 99%

Decellularization of pericardial tissue and its impact on tensile viscoelasticity and glycosaminoglycan content

et al. 2011

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“…Several combination ECM materials isolated from mammalian sources are commercially available [52,77] , while others are in various stages of development and characterization [19,[78][79][80] .…”

Section: Polymersmentioning

confidence: 99%

Biomaterials for spinal cord repair

Haggerty

Oudega

2013

Neurosci. Bull.

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Spinal cord injury (SCi) results in permanent loss of function leading to often devastating personal, economic and social problems. A contributing factor to the permanence of SCi is that damaged axons do not regenerate, which prevents the re-establishment of axonal circuits involved in function. Many groups are working to develop treatments that address the lack of axon regeneration after SCi. The emergence of biomaterials for regeneration and increased collaboration between engineers, basic and translational scientists, and clinicians hold promise for the development of effective therapies for SCi. A plethora of biomaterials is available and has been tested in various models of SCi. Considering the clinical relevance of contusion injuries, we primarily focus on polymers that meet the specific criteria for addressing this type of injury. Biomaterials may provide structural support and/or serve as a delivery vehicle for factors to arrest growth inhibition and promote axonal growth. Designing materials to address the specific needs of the damaged central nervous system is crucial and possible with current technology. Here, we review the most prominent materials, their optimal characteristics, and their potential roles in repairing and regenerating damaged axons following SCi.Keywords: spinal cord injury; axon regeneration; biodegradable materials; extracellular matrix proteins; functional recovery; growth factor; guidance; injury and repair; spinal motor neuron IntroductionSpinal cord injury (SCi) causes loss of neurons and axons resulting in motor and sensory function impairments. There are thousands of new cases of SCi in the world annually [1,2] occurring often in young adults. The lack of endogenous repair and the significant costs to the individual, family and society [1] are important motivations behind the continuing efforts to develop effective therapies. Promoting axonal regeneration is considered a potential repair strategy because it could lead to recovery of axonal circuits involved in motor and/or sensory function [3] .The central nervous system (CNS) neurons are intrinsically capable of some regeneration of damaged axons [4] but their attempts after SCi are frustrated by structural and chemical obstructions in the damaged nervous tissue [5] . Spinal cord repair approaches typically lead to small functional gains. one feature that most of these approaches have in common is a poor axonal regeneration response, suggesting that promoting axonal regeneration, including synapse formation, is essential to achieve substantial functional repair after SCi. if so, it is rational to anticipate that effective spinal cord therapies will need to include interventions addressing the axon growth-inhibition that leads to the poor axonal growth responses. As introduced above, axonal regeneration is considered an important repair mechanism for the injured spinal cord. Therefore, this review focuses on materials that have shown most promise to elicit an axonal regeneration response to foster functional restoration after ...

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“…In a randomized control trial of the Restore 1 patch, Iannotti et al [27] demonstrated no improvement in patients with Restore 1 augmentation when compared with patients who underwent the same procedure without xenograft addition. More recently, Restore 1 was found to contain a relatively high level of DNA within its matrix (1.13 ± 0.01 ng DNA/ mg dry weight) and has also resulted in an inflammatory result in 20% of patients whose rotator cuff tears were repaired with Restore 1 augmentation [22,38]. In a retrospective analysis, Walton et al [55] reported that patients whose rotator cuffs had been repaired with the Restore 1 patch experienced decreased post-repair strength, increased shoulder impingement, slower pain resolution after activity, and no decrease in retear rate when compared to patients whose rotator cuff tears had been repaired using standard surgical techniques.…”

Section: Ecm Augmentationmentioning

confidence: 99%

“…Zimmer 1 Collagen Repair Patch, marketed as Permacol 1 , (Tissue Science Laboratories PLC, Aldershot, Hampshire, UK), is a porcine dermal graft with no measurable DNA in its matrix and no reported inflammatory responses in patients [22]. One retrospective analysis of rotator cuff repair using Permacol 1 showed improved functional scores by 50% at 4.5 years postrepair [3].…”

Section: Ecm Augmentationmentioning

confidence: 99%

“…Allogenic ECMs can still contain some DNA from their allogenic source and may induce inflammatory responses in the host [22]. These inflammatory responses can cause pain and edema at the site of repair and may increase the degeneration of the rotator cuff repair that has been documented in the initial degenerative process of the rotator cuff [59].…”

Section: Ecm Augmentationmentioning

confidence: 99%

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Strategies in Biologic Augmentation of Rotator Cuff Repair: A Review

Cheung

Silverio

Sperling

2010

Clinical Orthopaedics &Amp; Related Research

147

103

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Background Degenerative rotator cuff tears are increasing with the aging population, and healing is not uniform after surgery. Rotator cuffs may show improved healing when biologic factors are added during surgery. Questions/purposes We asked: (1) What cellular processes are involved in normal bone-to-tendon healing? (2) What approaches are being developed in tendon augmentation? (3) What approaches are being developed with the addition of growth factors? Methods We reviewed research in relating to biologic augmentation and cellular processes involved in rotator cuff repair, focusing on animal models of rotator cuff repair and nonrandomized human trials. Results Regular bone-to-tendon healing forms a fibrous junction between tendon and bone that is distinct from the original bone-to-tendon junction. Tendon augmentation with cellular components serves as scaffolding for fibroblastic cells and a possible source of growth factors and fibroblastic cells. Extracellular matrices provide a scaffold for incoming fibroblastic cells, although current research does not conclusively confirm which if any of these scaffolds enhance repair owing in part to intermanufacturer variations and the limited human research. Growth factors and platelet-rich-plasma are established in other fields of research and may enhance repair but have not been rigorously tested. Conclusions There is potential application of biologic augmentation to improve healing after rotator cuff repair. However, research in this field is still inconclusive and has not been sufficiently demonstrated to merit regular clinical use. Future human trials can elucidate the use of biologic augmentation in rotator cuff repairs.

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Quantification of DNA in Biologic Scaffold Materials

Cited by 438 publications

References 42 publications

Decellularization of pericardial tissue and its impact on tensile viscoelasticity and glycosaminoglycan content

Decellularization of pericardial tissue and its impact on tensile viscoelasticity and glycosaminoglycan content

Biomaterials for spinal cord repair

Strategies in Biologic Augmentation of Rotator Cuff Repair: A Review

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