Healing Osteoarthritis: Engineered Proteins Created for Therapeutic Cartilage Regeneration
Millions of people worldwide are afflicted with painfulosteoarthritis, which is characterized by degradationof articular cartilage found in major joints such as thehip or knee. Symptoms include inflammation, pain,and decreased mobility. Because cartilage has a limitedability to self-heal, researcher...
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| Format: | Article |
| Language: | English |
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Purdue University Press
2012-01-01
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| Series: | Journal of Purdue Undergraduate Research |
| Online Access: | http://docs.lib.purdue.edu/cgi/viewcontent.cgi?article=1032&context=jpur |
| _version_ | 1818411216801890304 |
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| author | Kevin M. Cherry |
| author_facet | Kevin M. Cherry |
| author_sort | Kevin M. Cherry |
| collection | DOAJ |
| description | Millions of people worldwide are afflicted with painfulosteoarthritis, which is characterized by degradationof articular cartilage found in major joints such as thehip or knee. Symptoms include inflammation, pain,and decreased mobility. Because cartilage has a limitedability to self-heal, researchers have focused efforts onmethods that trigger cartilage regeneration. Our approachis to develop an injectable, protein-based hydrogel withmechanical properties analogous to healthy articularcartilage. The hydrogel provides an environment for cellgrowth and stimulates new tissue formation. We utilizedrecombinant DNA technology to create multifunctional,elastomeric proteins. The recombinant proteins weredesigned with biologically active domains to influence cellbehavior and resilin structural domains that mimic thestiffness of native cartilage. Resilin, a protein found in thewing and leg joints of mosquitoes, provided inspiration forthe mechanical domain in the recombinant protein. Thenew resilin-based protein was expressed in E. coli bacteria.Forming hydrogels requires a large quantity of engineeredprotein, so parameters such as bacterial host, incubationtemperature, expression time, and induction method wereoptimized to increase the protein yield. Using salt toprecipitate the protein and exploiting resilin’s heat stability,27 mg/L of recombinant protein was recovered at 95%purity. The protein expression and purification protocolswere established by analyzing experimental samples onSDS-PAGE gels and by Western blotting. The mechanicalproperties and interactions with stem cells are currentlybeing evaluated to assess the potential of the resilin-basedhydrogel as a treatment for osteoarthritis. |
| first_indexed | 2024-12-14T10:27:54Z |
| format | Article |
| id | doaj.art-429cfe3eef80440aad1c921599bdd801 |
| institution | Directory Open Access Journal |
| issn | 2158-4044 2158-4052 |
| language | English |
| last_indexed | 2024-12-14T10:27:54Z |
| publishDate | 2012-01-01 |
| publisher | Purdue University Press |
| record_format | Article |
| series | Journal of Purdue Undergraduate Research |
| spelling | doaj.art-429cfe3eef80440aad1c921599bdd8012022-12-21T23:06:16ZengPurdue University PressJournal of Purdue Undergraduate Research2158-40442158-40522012-01-0128182Healing Osteoarthritis: Engineered Proteins Created for Therapeutic Cartilage RegenerationKevin M. CherryMillions of people worldwide are afflicted with painfulosteoarthritis, which is characterized by degradationof articular cartilage found in major joints such as thehip or knee. Symptoms include inflammation, pain,and decreased mobility. Because cartilage has a limitedability to self-heal, researchers have focused efforts onmethods that trigger cartilage regeneration. Our approachis to develop an injectable, protein-based hydrogel withmechanical properties analogous to healthy articularcartilage. The hydrogel provides an environment for cellgrowth and stimulates new tissue formation. We utilizedrecombinant DNA technology to create multifunctional,elastomeric proteins. The recombinant proteins weredesigned with biologically active domains to influence cellbehavior and resilin structural domains that mimic thestiffness of native cartilage. Resilin, a protein found in thewing and leg joints of mosquitoes, provided inspiration forthe mechanical domain in the recombinant protein. Thenew resilin-based protein was expressed in E. coli bacteria.Forming hydrogels requires a large quantity of engineeredprotein, so parameters such as bacterial host, incubationtemperature, expression time, and induction method wereoptimized to increase the protein yield. Using salt toprecipitate the protein and exploiting resilin’s heat stability,27 mg/L of recombinant protein was recovered at 95%purity. The protein expression and purification protocolswere established by analyzing experimental samples onSDS-PAGE gels and by Western blotting. The mechanicalproperties and interactions with stem cells are currentlybeing evaluated to assess the potential of the resilin-basedhydrogel as a treatment for osteoarthritis.http://docs.lib.purdue.edu/cgi/viewcontent.cgi?article=1032&context=jpur |
| spellingShingle | Kevin M. Cherry Healing Osteoarthritis: Engineered Proteins Created for Therapeutic Cartilage Regeneration Journal of Purdue Undergraduate Research |
| title | Healing Osteoarthritis: Engineered Proteins Created for Therapeutic Cartilage Regeneration |
| title_full | Healing Osteoarthritis: Engineered Proteins Created for Therapeutic Cartilage Regeneration |
| title_fullStr | Healing Osteoarthritis: Engineered Proteins Created for Therapeutic Cartilage Regeneration |
| title_full_unstemmed | Healing Osteoarthritis: Engineered Proteins Created for Therapeutic Cartilage Regeneration |
| title_short | Healing Osteoarthritis: Engineered Proteins Created for Therapeutic Cartilage Regeneration |
| title_sort | healing osteoarthritis engineered proteins created for therapeutic cartilage regeneration |
| url | http://docs.lib.purdue.edu/cgi/viewcontent.cgi?article=1032&context=jpur |
| work_keys_str_mv | AT kevinmcherry healingosteoarthritisengineeredproteinscreatedfortherapeuticcartilageregeneration |