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Article ; Online: Fabrication of Size-Controlled and Emulsion-Free Chitosan-Genipin Microgels for Tissue Engineering Applications.

Stager, Michael A / Erickson, Christopher B / Payne, Karin A / Krebs, Melissa D

Journal of visualized experiments : JoVE

2022 , Issue 182

Abstract: Chitosan microgels are of significant interest in tissue engineering due to their wide range of applications, low cost, and immunogenicity. However, chitosan microgels are commonly fabricated using emulsion methods that require organic solvent rinses, ... ...

Abstract	Chitosan microgels are of significant interest in tissue engineering due to their wide range of applications, low cost, and immunogenicity. However, chitosan microgels are commonly fabricated using emulsion methods that require organic solvent rinses, which are toxic and harmful to the environment. The present protocol presents a rapid, non-cytotoxic, non-emulsion-based method for fabricating chitosan-genipin microgels without the need for organic solvent rinses. The microgels described herein can be fabricated with precise size control. They exhibit sustained release of biomolecules, making them highly relevant for tissue engineering, biomaterials, and regenerative medicine. Chitosan is crosslinked with genipin to form a hydrogel network, then passed through a syringe filter to produce the microgels. The microgels can be filtered to create a range of sizes, and they show pH-dependent swelling and degrade over time enzymatically. These microgels have been employed in a rat growth plate injury model and were demonstrated to promote increased cartilage tissue repair and to show complete degradation at 28 days in vivo. Due to their low cost, high convenience, and ease of fabrication with cytocompatible materials, these chitosan microgels present an exciting and unique technology in tissue engineering.
MeSH term(s)	Animals ; Chitosan ; Emulsions ; Iridoids ; Microgels ; Rats ; Solvents ; Tissue Engineering
Chemical Substances	Emulsions ; Iridoids ; Microgels ; Solvents ; Chitosan (9012-76-4) ; genipin (A3V2NE52YG)
Language	English
Publishing date	2022-04-13
Publishing country	United States
Document type	Journal Article ; Video-Audio Media ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/63857
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Fabrication of size-controlled and emulsion-free chitosan-genipin microgels for tissue engineering applications

Stager, Michael A. / Erickson, Christopher B. / Payne, Karin A. / Krebs, Melissa D.

Journal of visualized experiments. 2022 Apr. 13, , no. 182

2022

Abstract	Chitosan microgels are of significant interest in tissue engineering due to their wide range of applications, low cost, and immunogenicity. However, chitosan microgels are commonly fabricated using emulsion methods that require organic solvent rinses, which are toxic and harmful to the environment. The present protocol presents a rapid, non-cytotoxic, non-emulsion-based method for fabricating chitosan-genipin microgels without the need for organic solvent rinses. The microgels described herein can be fabricated with precise size control. They exhibit sustained release of biomolecules, making them highly relevant for tissue engineering, biomaterials, and regenerative medicine. Chitosan is crosslinked with genipin to form a hydrogel network, then passed through a syringe filter to produce the microgels. The microgels can be filtered to create a range of sizes, and they show pH-dependent swelling and degrade over time enzymatically. These microgels have been employed in a rat growth plate injury model and were demonstrated to promote increased cartilage tissue repair and to show complete degradation at 28 days in vivo. Due to their low cost, high convenience, and ease of fabrication with cytocompatible materials, these chitosan microgels present an exciting and unique technology in tissue engineering.
Keywords	biocompatible materials ; cartilage ; chitosan ; crosslinking ; emulsions ; growth plate ; hydrogels ; immunogenicity ; medicine ; microgels ; models ; pH ; rats ; solvents ; syringes ; tissue repair ; toxicity
Language	English
Dates of publication	2022-0413
Size	p. e63857.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/63857
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: A timeseries analysis of the fracture callus extracellular matrix proteome during bone fracture healing.

Erickson, Christopher B / Hill, Ryan / Pascablo, Donna / Kazakia, Galateia / Hansen, Kirk / Bahney, Chelsea

Journal of life sciences (Westlake Village, Calif.)

2022 Volume 3, Issue 4, Page(s) 1–30

Abstract: While most bones fully self-heal, certain diseases require bone allograft to assist with fracture healing. Bone allografts offer promise as treatments for such fractures due to their osteogenic properties. However, current bone allografts made of ... ...

Abstract	While most bones fully self-heal, certain diseases require bone allograft to assist with fracture healing. Bone allografts offer promise as treatments for such fractures due to their osteogenic properties. However, current bone allografts made of decellularized bone extracellular matrix (ECM) have high failure rates, and thus grafts which improve fracture healing outcomes are needed. Understanding specific changes to the ECM proteome during normal fracture healing would enable the identification of key proteins that could be used enhance osteogenicity of bone allograft. Here, we performed a timeseries analysis of the fracture callus in mice to investigate proteomic and mineralization changes to the ECM at key stages of fracture healing. We found that changes to the ECM proteome largely coincide with the distinct phases of fracture healing. Basement membrane proteins (AGRN, COL4, LAMA), cartilage proteins (COL2A1, ACAN), and collagen crosslinking enzymes (LOXL, PLOD, ITIH) were initially upregulated, followed by bone specific proteoglycans and collagens (IBSP, COL1A1). Various tissue proteases (MMP2, 9, 13, 14; CTSK, CTSG, ELANE) were expressed at different levels throughout fracture healing. These changes coordinated with mineralization of the fracture callus, which increased steeply during the initial stages of healing. Interestingly the later timepoint was characterized by a response to wound healing and high expression of clotting factors (F2, 7, 9, 10). We identified ELANE and ITIH2 as tissue remodeling enzymes having no prior known involvement with fracture healing. This data can be further mined to identify regenerative proteins for enhanced bone graft design.
Language	English
Publishing date	2022-06-24
Publishing country	United States
Document type	Journal Article
ISSN	2688-1020
ISSN (online)	2688-1020
DOI	10.36069/JoLS/20220601
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Article: Analysis of Physeal Fractures from the United States National Trauma Data Bank.

Fuchs, Joseph R / Gibly, Romie F / Erickson, Christopher B / Thomas, Stacey M / Hadley Miller, Nancy / Payne, Karin A

Children (Basel, Switzerland)

2022 Volume 9, Issue 6

Abstract: Background: Pediatric long-bone physeal fractures can lead to growth deformities. Previous studies have reported that physeal fractures make up 18-30% of total fractures. This study aimed to characterize physeal fractures with respect to sex, age, ... ...

Abstract	Background: Pediatric long-bone physeal fractures can lead to growth deformities. Previous studies have reported that physeal fractures make up 18-30% of total fractures. This study aimed to characterize physeal fractures with respect to sex, age, anatomic location, and Salter-Harris (SH) classification from a current multicenter national database. Methods: A retrospective cohort study was performed using the 2016 United States National Trauma Data Bank (NTDB). Patients ≤ 18 years of age with a fracture of the humerus, radius, ulna, femur, tibia, or fibula were included. Results: The NTDB captured 132,018 patients and 58,015 total fractures. Physeal fractures made up 5.7% (3291) of all long-bone fractures, with males accounting for 71.0% (2338). Lower extremity physeal injuries comprised 58.6% (1929) of all physeal fractures. The most common site of physeal injury was the tibia comprising 31.8% (1047), 73.9% (774) of which were distal tibia fractures. Physeal fractures were greatest at 11 years of age for females and 14 years of age for males. Most fractures were SH Type II fractures. Discussion and conclusions: Our analysis indicates that 5.7% of pediatric long-bone fractures involved the physis, with the distal tibia being the most common. These findings suggest a lower incidence of physeal fractures than previous studies and warrant further investigation.
Language	English
Publishing date	2022-06-18
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2732685-8
ISSN	2227-9067
ISSN	2227-9067
DOI	10.3390/children9060914
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Article: ECM-Focused Proteomic Analysis of Ear Punch Regeneration in

McCabe, Maxwell C / Okamura, Daryl M / Erickson, Christopher B / Perry, Blair W / Brewer, Chris M / Nguyen, Elizabeth D / Saviola, Anthony J / Majesky, Mark W / Hansen, Kirk C

bioRxiv : the preprint server for biology

2023

Abstract: In mammals, significant injury is generally followed by the formation of a fibrotic scar which provides structural integrity but fails to functionally restore damaged tissue. Spiny mice of the ... ...

Abstract	In mammals, significant injury is generally followed by the formation of a fibrotic scar which provides structural integrity but fails to functionally restore damaged tissue. Spiny mice of the genus
Language	English
Publishing date	2023-10-17
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.10.11.561940
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Article: Trans-Omics analysis of post injury thrombo-inflammation identifies endotypes and trajectories in trauma patients.

Cohen, Mitchell J / Erickson, Christopher B / Lacroix, Ian S / Debot, Margaret / Dzieciatkowska, Monika / Schaid, Terry R / Hallas, Morgan W / Thielen, Otto N / Cralley, Alexis L / Banerjee, Anirban / Moore, Ernest E / Silliman, Christopher C / D'Alessandro, Angelo / Hansen, Kirk C

bioRxiv : the preprint server for biology

2023

Abstract: Understanding and managing the complexity of trauma-induced thrombo-inflammation necessitates an innovative, data-driven approach. This study leveraged a trans-omics analysis of longitudinal samples from trauma patients to illuminate molecular endotypes ... ...

Abstract	Understanding and managing the complexity of trauma-induced thrombo-inflammation necessitates an innovative, data-driven approach. This study leveraged a trans-omics analysis of longitudinal samples from trauma patients to illuminate molecular endotypes and trajectories that underpin patient outcomes, transcending traditional demographic and physiological characterizations. We hypothesize that trans-omics profiling reveals underlying clinical differences in severely injured patients that may present with similar clinical characteristics but ultimately have very different responses to treatment and clinical outcomes. Here we used proteomics and metabolomics to profile 759 of longitudinal plasma samples from 118 patients at 11 time points and 97 control subjects. Results were used to define distinct patient states through data reduction techniques. The patient groups were stratified based on their shock severity and injury severity score, revealing a spectrum of responses to trauma and treatment that are fundamentally tied to their unique underlying biology. Ensemble models were then employed, demonstrating the predictive power of these molecular signatures with area under the receiver operating curves of 80 to 94% for key outcomes such as INR, ICU-free days, ventilator-free days, acute lung injury, massive transfusion, and death. The molecularly defined endotypes and trajectories provide an unprecedented lens to understand and potentially guide trauma patient management, opening a path towards precision medicine. This strategy presents a transformative framework that aligns with our understanding that trauma patients, despite similar clinical presentations, might harbor vastly different biological responses and outcomes.
Language	English
Publishing date	2023-09-09
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.08.16.553446
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Article ; Online: Blockade of Osteoclast-Mediated Bone Resorption With a RANKL-Inhibitor Enhances Bone Formation in a Rat Spinal Fusion Model.

Payne, Karin A / Shaw, Nichole M / Erickson, Christopher B / Yarger, Peter / Yu, Yangyi / Baldini, Todd / Kleck, Christopher J / Patel, Vikas V / Burger, Evalina L

Spine

2022 Volume 47, Issue 16, Page(s) 1165–1171

Abstract: Study design: Rat spine fusion model.: Objective: The present study aimed to determine whether administration of osteoprotegerin (OPG) in a rat model of spinal fusion increases bone volume, bone density, and decreases osteoclasts in the fusion mass.!# ...

Abstract	Study design: Rat spine fusion model. Objective: The present study aimed to determine whether administration of osteoprotegerin (OPG) in a rat model of spinal fusion increases bone volume, bone density, and decreases osteoclasts in the fusion mass. Summary of background data: OPG is a soluble RANK-ligand inhibitor that blocks osteoclast differentiation and activation. This makes it a potential agent to control the remodeling process and enhance bone mass during spinal fusion. Materials and methods: Forty-eight male Sprague-Dawley rats received a one-level spinal fusion of L4-L5 with bone allograft. Rats were then divided into four groups according to initiation of treatment: (1) saline on day 0 (saline), (2) OPG on day 0 (OPG D0), (3) OPG on day 10 (OPG D10), and (4) OPG on day 21 (OPG D21) postsurgery. After their initial injection, rats received weekly subcutaneous injections of OPG (10 mg/kg) and were euthanized six weeks postsurgery. MicroCT analysis of the fusion site and histological analysis of bone surface for quantification of osteoclast lining was performed. Results: Increased bone volume in the fusion site and around the spinous process was seen in OPG D0 and OPG D10 when compared with saline. Mean trabecular thickness was greater in all groups receiving OPG compared with saline, with OPG D0 and OPG D10 having significantly greater mean trabecular thickness than OPG D21. All OPG groups had less bone surface lined with osteoclasts when compared with Saline, with OPG D0 and OPG D10 having fewer than OPG D21. Conclusions: This study indicates that OPG inhibited osteoclast bone resorption, which led to greater bone at the fusion site. Future studies investigating OPG on its own or in combination with an osteogenic factor to improve spinal fusion outcomes are warranted to further elucidate its potential therapeutic effect.
MeSH term(s)	Animals ; Bone Resorption/drug therapy ; Bone Resorption/pathology ; Male ; Osteoclasts ; Osteogenesis ; Osteoprotegerin ; RANK Ligand/pharmacology ; RANK Ligand/therapeutic use ; Rats ; Rats, Sprague-Dawley ; Spinal Fusion
Chemical Substances	Osteoprotegerin ; RANK Ligand
Language	English
Publishing date	2022-07-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	752024-4
ISSN	1528-1159 ; 0362-2436
ISSN (online)	1528-1159
ISSN	0362-2436
DOI	10.1097/BRS.0000000000004412
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Article: A rat tibial growth plate injury model to characterize repair mechanisms and evaluate growth plate regeneration strategies

Erickson, Christopher B / Shaw, Nichole / Hadley-Miller, Nancy / Riederer, Michael S / Krebs, Melissa D / Payne, Karin A

Journal of visualized experiments. 2017 July 04, , no. 125

2017

Abstract: A third of all pediatric fractures involve the growth plate and can result in impaired bone growth. The growth plate (or physis) is cartilage tissue found at the end of all long bones in children that is responsible for longitudinal bone growth. Once ... ...

Abstract	A third of all pediatric fractures involve the growth plate and can result in impaired bone growth. The growth plate (or physis) is cartilage tissue found at the end of all long bones in children that is responsible for longitudinal bone growth. Once damaged, cartilage tissue within the growth plate can undergo premature ossification and lead to unwanted bony repair tissue, which forms a "bony bar." In some cases, this bony bar can result in bone growth deformities, such as angular deformities, or it can completely halt longitudinal bone growth. There is currently no clinical treatment that can fully repair an injured growth plate. Using an animal model of growth plate injury to better understand the mechanisms underlying bony bar formation and to identify ways to inhibit it is a great opportunity to develop better treatments for growth plate injuries. This protocol describes how to disrupt the rat proximal tibial growth plate using a drill-hole defect. This small animal model reliably produces a bony bar and can result in growth deformities similar to those seen in children. This model allows for investigation into the molecular mechanisms of bony bar formation and serves as a means to test potential treatment options for growth plate injuries.
Keywords	animal models ; bone formation ; cartilage ; children ; growth plate ; rats ; tibia
Language	English
Dates of publication	2017-0704
Size	p. e55571.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/55571
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Article ; Online: In vivo degradation rate of alginate-chitosan hydrogels influences tissue repair following physeal injury.

Erickson, Christopher B / Newsom, Jake P / Fletcher, Nathan A / Feuer, Zachary M / Yu, Yangyi / Rodriguez-Fontan, Francisco / Hadley Miller, Nancy / Krebs, Melissa D / Payne, Karin A

Journal of biomedical materials research. Part B, Applied biomaterials

2020 Volume 108, Issue 6, Page(s) 2484–2494

Abstract: The physis is a cartilaginous tissue in children's long bones that is responsible for bone elongation. Physeal injuries can heal with bony repair tissue known as a "bony bar," and this can cause growth deformities. Current treatments involve surgical ... ...

Abstract	The physis is a cartilaginous tissue in children's long bones that is responsible for bone elongation. Physeal injuries can heal with bony repair tissue known as a "bony bar," and this can cause growth deformities. Current treatments involve surgical resection of the bony bar and insertion of inert materials in hopes of preventing bony bar re-formation and preserving bone elongation. However, these materials frequently fail and the bony bar commonly returns. This study investigated alginate-chitosan hydrogels as interpositional materials to block bony bar formation in a rat model of physeal injury. Further, biomaterial properties such as substrate stiffness, permeability, and degradation rate were studied. Different ratio alginate:chitosan hydrogels with or without calcium cross-linking were tested for their inhibition of bony bar formation and restoration of the injured physis. Alginate:chitosan were mixed (a) 90:10 with calcium (90:10 + Ca); (b) 50:50 with calcium (50:50 + Ca); (c) 50:50 without calcium (50:50 - Ca); and (d) 50:50 made with irradiated alginate (IA) and without calcium. We found that repair tissue was determined primarily by the in vivo degradation rate of alginate-chitosan hydrogels. 90:10 + Ca had a slow degradation rate, prevented cellular infiltration, and produced the most bony bar tissue while having softer, more permeable material properties. IA had the fastest degradation, showed high cellular infiltration, and produced the most cartilage-like tissue while having stiffer, less permeable material properties. Our results suggest that the in vivo biomaterial degradation rate is a dynamic property that can be optimized to influence cell fate and tissue repair in physeal injuries.
MeSH term(s)	Alginates/metabolism ; Animals ; Biocompatible Materials/metabolism ; Calcium/chemistry ; Calcium/pharmacology ; Chitosan/metabolism ; Cross-Linking Reagents ; Growth Plate/growth & development ; Growth Plate/pathology ; Hydrogels ; Mechanical Phenomena ; Osteogenesis ; Permeability ; Rats ; Rats, Sprague-Dawley ; Rheology ; Wound Healing
Chemical Substances	Alginates ; Biocompatible Materials ; Cross-Linking Reagents ; Hydrogels ; Chitosan (9012-76-4) ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2020-02-08
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2099992-6
ISSN	1552-4981 ; 1552-4973 ; 0021-9304
ISSN (online)	1552-4981
ISSN	1552-4973 ; 0021-9304
DOI	10.1002/jbm.b.34580
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Zs.A 6196: Show issues

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Article ; Online: Anti-VEGF antibody delivered locally reduces bony bar formation following physeal injury in rats.

Erickson, Christopher B / Newsom, Jake P / Fletcher, Nathan A / Yu, Yangyi / Rodriguez-Fontan, Francisco / Weatherford, Shane A / Hadley-Miller, Nancy / Krebs, Melissa D / Payne, Karin A

Journal of orthopaedic research : official publication of the Orthopaedic Research Society

2020 Volume 39, Issue 8, Page(s) 1658–1668

Abstract: Physeal injuries can result in the formation of a "bony bar" which can lead to bone growth arrest and deformities in children. Vascular endothelial growth factor (VEGF) has been shown to play a role in bony bar formation, making it a potential target to ... ...

Abstract	Physeal injuries can result in the formation of a "bony bar" which can lead to bone growth arrest and deformities in children. Vascular endothelial growth factor (VEGF) has been shown to play a role in bony bar formation, making it a potential target to inhibit bony repair tissue after physeal injury. The goal of this study was to investigate whether the local delivery of anti-VEGF antibody (α-VEGF; 7.5 μg) from alginate:chitosan hydrogels to the tibial physeal injury site in rats prevents bony bar formation. We tested the effects of quick or delayed delivery of α-VEGF using both 90:10 and 50:50 ratio alginate:chitosan hydrogels, respectively. Male and female 6-week-old Sprague-Dawley rats received a tibial physeal injury and the injured site injected with alginate-chitosan hydrogels: (1) 90:10 (Quick Release); (2) 90:10 + α-VEGF (Quick Release + α-VEGF); (3) 50:50 (Slow Release); (4) 50:50 + α-VEGF (Slow Release + α-VEGF); or (5) Untreated. At 2, 4, and 24 weeks postinjury, animals were euthanized and tibiae assessed for bony bar and vessel formation, repair tissue type, and limb lengthening. Our results indicate that Quick Release + α-VEGF reduced bony bar and vessel formation, while also increasing cartilage repair tissue. Further, the quick release of α-VEGF neither affected limb lengthening nor caused deleterious side-effects in the adjacent, uninjured physis. This α-VEGF treatment, which inhibits bony bar formation without interfering with normal bone elongation, could have positive implications for children suffering from physeal injuries.
MeSH term(s)	Alginates ; Animals ; Antibodies/immunology ; Chitosan ; Female ; Growth Plate/metabolism ; Hydrogels ; Male ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A/metabolism
Chemical Substances	Alginates ; Antibodies ; Hydrogels ; Vascular Endothelial Growth Factor A ; Chitosan (9012-76-4)
Language	English
Publishing date	2020-11-24
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	605542-4
ISSN	1554-527X ; 0736-0266
ISSN (online)	1554-527X
ISSN	0736-0266
DOI	10.1002/jor.24907
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