Cell attachment coatings and methods using phosphorous-containing photoreagent
View Patent ↗Cell attachment coatings for articles such as implantable medical devices and cell culture vessels are disclosed. The coatings include an intermediate coater layer which includes a phosphorous-containing component that is bonded in the coating by reacted photoreactive functional groups. The coating also include a second coated layer including an immobilized ECM protein or peptide that includes an active portion of an ECM protein that is able to serve as an outer layer to contact cells during use. The coatings promoted enhanced cell binding and growth.
1. An article comprising a biocompatible cell attachment coating comprising:
(a) an intermediate coated layer comprising a first component that is a compound of one of Formulas I, II, or III
wherein, in Formula I and II, Photo 1 and Photo 2 are, independently, photoreactive groups comprising an aryl ketone group, and at least one covalent linkage between Photo 1 and P, Photo 2 and P, or both, is interrupted by at least one heteroatom, and R is alkyl, aryl, a photoreactive group, hydroxyl or salt thereof;
wherein, in Formula III, Photo 1 and Photo 2 are, independently, photoreactive groups comprising an aryl ketone functional group, and at least one covalent linkage between Photo 1 and P, Photo 2 and P, or both, is interrupted by at least one heteroatom; Y represents a covalent bond, N, O, linear or branched C 1 -C 10 alkyl; and R 1 and R 2 are independently is alkyl, aryl, a photoreactive group, hydroxyl or salt thereof;
wherein the first component is immobilized in the coating via the photoreactive group; and
(b) a second coated layer comprising a cell attachment molecule comprising an extracellular matrix (ECM) protein, or a peptide that includes an active portion of an ECM protein that is immobilized in the coating;
wherein the intermediate coated layer is positioned between the second coated layer and a surface of the article.
2. The article of claim 1 wherein the extracellular matrix protein is selected from the group consisting of fibronectin, laminin, collagen, procollagen, elastin, vitronectin, tenascin, entactin, fibrinogen, thrombospondin, osteopontin (bone sialoprotein), osteocalcin, von Willebrand Factor, or the peptide includes an active portion of the recited ECM proteins for cell attachment.
3. The article of claim 2 comprising collagen or laminin, or a peptide comprising a RGD motif.
4. The article of claim 3 wherein the peptide comprising a RGD motif is selected from the group consisting of RGD, RGDS (SEQ ID NO:14), RGDT (SEQ ID NO:15), GRGD (SEQ ID NO:16), GRGDS (SEQ ID NO:17), GRGDG (SEQ ID NO:18), GRGDSP (SEQ ID NO:13), GRGDSG (SEQ ID NO:19), GRGDNP (SEQ ID NO:20), GRGDSPK (SEQ ID NO:21), GRGDSY (SEQ ID NO:22), YRGDS (SEQ ID NO:23), YRGDG (SEQ ID NO:24), YGRGD (SEQ ID NO:25), CGRGDSY (SEQ ID NO:26), CGRGDSPK (SEQ ID NO:27), YAVTGRGDS (SEQ ID NO:28), RGDSPASSKP (SEQ ID NO:29), GRGDSPASSKG (SEQ ID NO:30), GCGYGRGDSPG (SEQ ID NO:31), GGGPHSRNGGGGGGRGDG (SEQ ID NO:32), RGDV (SEQ ID NO:33), RGDF (SEQ ID NO:34), GRGDF (SEQ ID NO:35), GRGDY (SEQ ID NO:36), GRGDVY (SEQ ID NO:37), and GRGDYPC (SEQ ID NO:38).
5. The article of claim 2 comprising type I collagen.
6. The article of claim 1 wherein the first component comprises a compound of Formula I.
7. The article of claim 1 wherein the first component is selected from the group consisting of:
8. The article of claim 7 wherein the first component is:
9. The article of claim 1 , wherein the article is a medical article and the photoreactive group is bonded to the surface of the medical article, to another first component in the coated layer, or both.
10. The article of claim 1 , wherein the intermediate coated layer has a thickness in the range of 10 nm to 100 nm.
11. The article of claim 1 , wherein the cell attachment molecule is covalently bonded to a target moiety selected from the first component, another matrix polypeptide, or both the first component and another matrix polypeptide.
12. The article of claim 1 , wherein the cell attachment molecule comprises a pendent reacted aryl ketone group allowing the cell attachment molecule to be immobilized in the coating.
13. The article of claim 1 , where the second layer further comprises a synthetic polymer, wherein the cell attachment molecule is bonded to the synthetic polymer, the synthetic polymer further comprising a pendent reacted aryl ketone group allowing the cell attachment molecule to be immobilized in the coating.
14. The article of claim 1 , wherein the coating comprises a mole to weight ratio of phosphorous-containing groups to the cell attachment molecule in the range of about 0.5 mmol/g to about 100 mmol/g.
15. The article of claim 1 , which is a medical article that is insertable or implantable into a portion of the body.
16. The article of claim 1 , wherein the article is a cell culture article.
17. A method for forming a coated article comprising steps of:
(a) applying a first composition on an article surface, the first composition comprising first component that is a compound of one of Formulas I, II, or III
wherein, in Formula I and II, Photo 1 and Photo 2 are, independently, photoreactive groups comprising an aryl ketone group, and at least one covalent linkage between Photo 1 and P, Photo 2 and P, or both, is interrupted by at least one heteroatom, and R is alkyl, aryl, a photoreactive group, hydroxyl or salt thereof;
wherein, in-Formula III, Photo 1 and Photo 2 are, independently, photoreactive groups comprising an aryl ketone group, and at least one covalent linkage between Photo 1 and P, Photo 2 and P, or both, is interrupted by at least one heteroatom; Y represents a covalent bond, N, O, linear or branched C 1 -C 10 alkyl; and R 1 and R 2 are independently is alkyl, aryl, a photoreactive group, hydroxyl or salt thereof;
(b) providing a second composition on the article surface, the second composition comprising a cell attachment molecule comprising an ECM protein, or a peptide that includes an active portion of an ECM protein for cell attachment;
(c) irradiating the device surface to cause bonding of the aryl functional group to a target moiety, wherein irradiating is performed after step (a), after step (b), or both after steps (a) and (b).
18. A method for promoting attachment and proliferation of cells to an article surface, the method comprising steps of
(a) providing an article of claim 1 ; and
(b) placing the article in an environment comprising cells, wherein the cells become attached to the coating.
19. The method of claim 18 where the article is an implantable or insertable medical article and the step of placing comprises implanting or inserting the article in the body.
20. The method of claim 18 where the article in a cell culture article and the step of placing comprises placing cells in the article in vitro.