Information for the AlignCol™ Products

Question:
What is the amount of collagen deposited per slide (8 X 15 mm) on the AlignCol™ products?
Answer:
The approximate amount of the collagen per one glass slide of AlignCol™ product is 0.5 mg.

Question:
What is the average size of each collagen fibril deposited on the glass slide of the AlignCol product?
Answer:
1) The typical AlignCol™ Braided product has collagen fibrils with a diameter in the range of 20 – 100 nm.
2) The typical AlignCol™ Crimped product has collagen fibrils with a diameter in the range of 100 – 300 nm.

Osmolality and Conductivity of Advanced BioMatrix’s Collagen Products

Osmolality and Conductivity of Advanced BioMatrix’s Collagen Products

Collagen Name	Source	Telo-Peptide Intact	Approximate Concentration	Average Osmolality (mOsm)	Average Conductivity (µS/cm)
PureCol®	Bovine	No	3 mg/ml	21	2328
Nutragen®	Bovine	No	6 mg/ml	22	2635
VitroCol®	Human	No	3 mg/ml	19	2153
TeloCol™	Bovine	Yes	6 mg/ml	12	2230
Rat Tail	Rat	Yes	4 mg/ml	20	300

Degradation of PureCol Collagen Matrices In Vivo

Question: Is there any published data measuring the time it takes to completely degrade PureCol collagen matrices in vivo?

 Answer:  There has been some experimental research conducted on degradation rates of PureCol collagen matrices.  See an abstract  titled, ‘Preparation of [3H]collagen for studies of the biologic fate of xenogenic collagen implants in vivo’  McPherson JM, Sawamura SJ, Conti A., J Invest Dermatol. 1986 Jun;86(6):673-7, http://www.ncbi.nlm.nih.gov/pubmed/3711680.  See abstract below:

Abstract

Reduction of a commercially available, pepsin-solubilized, bovine dermal collagen (Vitrogen 100) (PureCol’s old product name) with sodium [3H]borohydride provided radiolabeled collagen preparations with specific activities ranging from 7.1-12.0 muCi/mg collagen. These specific activities were 2-3 times greater than those obtained by reduction of intact rat tail tendon collagen under similar conditions. The alpha, beta, and higher aggregate components of type I collagen were radiolabeled as well as the alpha component of a small amount of type III collagen present in the samples. Fractionation of cyanogen bromide peptides showed that alpha 1(I)CB7, alpha 1(I)CB8, and alpha 2(I)CB3,5 were the predominant peptides labeled by this procedure. Amino acid analysis indicated that the majority of the radioactivity was in reducible cross-links, precursors of these cross-links, and in hexosyllysine residues. Reconstitution experiments comparing this radiolabeled collagen with nonlabeled collagen showed them to be indistinguishable. Bacterial collagenase digestion of this reconstituted fibrillar collagen in both a lightly cross-linked (glutaraldehyde 0.0075%) and noncross-linked form provided evidence that digestion of labeled and nonlabeled collagens proceeded at similar rates. Thus, labeling did not change the properties of the collagen. Cross-linking made the preparation refractory to proteolytic degradation. Injection of fibrillar collagen preparations, spiked with radiolabeled collagen, into the guinea pig dermis followed by quantitation of the amount of radioactivity recovered from implant sites as a function of time, indicated that the lightly cross-linked samples also were more resistant to degradation in vivo than the noncross-linked preparation. The half-life of noncross-linked collagen was about 4 days while that of the cross-linked collagen was about 25 days. These degradation rates were much faster than observed for similar, nonlabeled samples injected into the dermis of humans, presumably due to a higher metabolic activity in the guinea pig dermis.

Staining of PureCol Collagen for Visualization by Confocal Imaging

 

Question: To visualize the collagen component in PureCol collagen scaffold, can the collagen be stained with an anti-bovine collagen antibody for confocal imaging? Since PureCol is an atelo-collagen can an anti-bovine collagen antibody be used?

Answer: Since the collagen in PureCol collagen contains approximately 95% Type I bovine collagen and 5% Type III bovine collagen, an anti-bovine collagen Type I antibody for your study can be used.

Comparison of Acid Soluble and Pepsin Soluble Collagen atelo vs telo

COMPARISON ACID SOLUBLE AND PEPSIN SOLUBLE COLLAGEN

N-Terminal Amino Acid Sequence of Bovine Type I Collagen:

Collagen	Aminoterminal Telopeptide	Triple Helical Region	Collagen Type I Polypeptide
	pQLSYGYDEKSTGISVP	GPMGPSGPRGLOGPOGAOG…….	α-1
Acid Soluble Collagen	pQLSYGYDEKSTGISVP	GPMGPSGPRGLOGPOGAOG…….	α-1
	pQFDAKGGGP	GPMGLMGPRGPOGASGOAG……	α-2
	ISVP	GPMGPSGPRGLOGPOGAOG…….	α-1
Pepsin Soluble Collagen	DEKSTGISVP	GPMGPSGPRGLOGPOGAOG…….	α-1
	DAKGGGP	GPMGLMGPRGPOGASGAOG……	α-2

Comparison of Collagen Gels:

Collagen	Telo-Peptide	Gel Strength	Gel Structure	Gel Appearance
Acid Soluble Collagen	Telo-peptide Intact	Slightly stronger than pepsin solubilized	Straight fibrils, mesh-like structure	Transparent to White Gel
Pepsin Soluble Collagen	Telo-peptide Removed	Slightly weaker than acid solubilized	Loosely twisted fibrils, smaller mesh work, sponge-like	Clear to Transparent Gel

Journal of Biological Chemistry, Vol. 275, no. 33 pp. 25870-25875, 2000

Domains in Tropoelastin That Mediate Elastin Deposition in Vitro and in Vivo*

Domains in Tropoelastin That Mediate Elastin Deposition in Vitro and in Vivo — Kozel et al 278 (20)_ 18491 — Journal of Biological Chemistry

PureCol and Cross-Linked Hyaluronan-Gelatin Tested for Rheological Properties for Tissue Engineering

Vanderhooft Rheology Analysis Final print form 2009 (2)

PEPTITE-2000 (RGD Peptide) Molecular Weight

Question: What is the molecular weight (MW) of PEPTITE-2000 (RGD Peptide)?

Answer: The PEPTITE-2000 has an average MW of 2334.8.

Dissolving PEPTITE-2000 RGD Peptide For Conjugation to a Lysine Residue

Question:  To conjugate PEPTITE-2000 RGD peptide to the NHS- group via the NH2 group on Lysine residue, what solvent is recommended since water or ethanol cannot be used?

Answer:  It is recommend that you use Dimethlsulfoxide (DMSO) or dichloromethane (DCM) to solubilize PEPTITE-2000 for this application.

Modified Dendrimer Cross-Linked Collagen-Based Matrices Using PureCol®

J Biomater Sci Polym Ed. 2011 Dec 2.

Modified Dendrimer Cross-Linked Collagen-Based Matrices Using PureCol.

Princz MA, Sheardown H.

Abstract

Dendrimer cross-linking has been achieved with pepsin digested over 80% type-I bovine collagen to create strong hydrogels with good cell compatibility. Herein we investigate the use of commercially available collagen-based products with the dendrimer cross-linking technology. Specifically PureCol(®) (PC), a 97% bovine type-I collagen, human collagen (HC) and human extracellular matrix (hECM) were concentrated, and then cross-linked with polypropyleneimine octaamine generation two dendrimers using 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) chemistry. PC gels with 30 and 20 mg/ml bovine collagen were fabricated, and despite similar concentrations to >80% type-I bovine collagen dendrimer cross-linked gels (CG), PC gels demonstrated increased swelling and decreased stability, as determined with collagenase digestion. The highly purified bovine (PC) and human sourced-collagen (HC) gels were similar in performance, but not as stable as the CG gels, which may correlate to the manufacturer’s collagen purification and storage. Finally, the addition of hECM components to PC to create PC-hECM gels, resulted in a looser gel network, compared to heparinized dendrimer cross-linked bovine >80% type-I collagen gels (CHG). However, all collagen-based gels supported 3T3 fibroblast cell growth over 4 days, indicating these gels may be suitable for tissue-engineering applications.