US-12622999-B2 - PCL patch tissue regeneration scaffold and method for manufacturing same

US12622999B2US 12622999 B2US12622999 B2US 12622999B2US-12622999-B2

Abstract

Disclosed is a method of treating a chronic tympanic perforation with a PCL patch tissue regeneration scaffold comprising preparing solution by adding polycaprolactone (PCL) and an acid to an organic solvent; preparing an electrospinning solution by adding a growth factor to the solution and stirring; collecting nanofibers arranged in a spindle shape on a collector by connecting the electrospinning solution prepared to a syringe pump and operating the electrospinning device; and administering the PCL patch tissue regeneration scaffold to a subject.

Inventors

Yun-Hoon Choung
Jong Hoon Chung
Hoon SEONWOO
Kyoung-Je JANG
Beomyong SHIN

Assignees

AJOU UNIVERSITY INDUSTRY-ACADEMIC COOPERATION FOUNDATION

Dates

Publication Date: 20260512
Application Date: 20230921
Priority Date: 20160712

Claims (2)

1 . A method of treating a chronic tympanic membrane perforation in a subject, comprising: preparing a spindle-shaped polycaprolactone (PCL) patch tissue regeneration scaffold by: preparing an electrospinning solution by adding the PCL and acetic acid to an organic solvent and stirring, wherein a concentration of the PCL in the organic solvent is 70 to 90 mg/ml; adding epithelial growth factor (EGF) and insulin growth factor binding protein (IGFBP) to the electrospinning solution and stirring at a temperature between 0° C. and 10° C., wherein the electrospinning solution comprises 0.16 to 16 μg/ml of the EGF and 0.16 to 16 μg/mL of the IGFBP; and collecting nanofibers on a cylindrical collector housing a single centrally located needle with a sharp end by electrospinning the electrospinning solution through a syringe pump at a flow rate of 0.2 to 3.0 μL/hr while applying 3 voltage of 18 kV across a distance of 10 cm between the single centrally located needle and the cylindrical collector, wherein the collected nanofibers are arranged in a radially aligned pattern converging from a periphery toward the single centrally located needle, thereby forming the spindle-shaped PCL patch tissue regeneration scaffold; and applying the spindle-shaped PCL patch tissue regeneration scaffold to the subject's tympanic membrane perforation of the subject without surgery, whereby the tympanic membrane at the chronic tympanie perforation is regenerated in the subject using the spindle-shaped PCL patch tissue regeneration scaffold loaded with the EGF and the IGFBP.
2 . A method of treating a chronic tympanic membrane perforation in a subject, comprising: preparing a spindle-shaped PCL-PEG patch tissue regeneration scaffold by: preparing an electrospinning solution by adding polycaprolactone (PCL), polyethylene glycol (PEG), and acetic acid to an organic solvent and stirring to dissolve the polymers, wherein the electrospinning solution comprises PCL at a concentration of 100 to 160 mg/ml and PEG at a concentration of 40 to 100 mg/ml; adding epithelial growth factor (EGF) and insulin growth factor binding protein (IGFBP) to the electrospinning solution and stirring at a temperature between 0° C. and 10° C., wherein the electrospinning solution comprises 0.16 to 16 μg/ml of the EGF and 0.16 to 16 μg/ml of the IGFBP; and collecting nanofibers on a cylindrical collector housing a single centrally located needle with a sharp end by electrospinning the electrospinning solution through a syringe pump at a flow rate of 0.2 to 3.0 μL/hr while applying a voltage of 18 k kV across a distance of 10 cm between the single centrally located needle and the cylindrical collector, wherein the collected nanofibers are arranged in a radially aligned pattern converging from a periphery toward the single centrally located needle, thereby forming the spindle-shaped PCL-PEG patch tissue regeneration scaffold; and applying the spindle-shaped PCL-PEG patch tissue regeneration scaffold to the tympanic membrane perforation of the subject without surgery; whereby the tympanic membrane perforation is regenerated in the subject using the spindle-shaped PCL-PEG patch tissue regeneration scaffold loaded with the EGF and the IGFBP.

Description

CROSS REFERENCE TO PRIOR APPLICATIONS This application is a Continuation Application of U.S. patent application Ser. No. 16/317,028 filed Mar. 25, 2019, which is a National Stage Application of PCT International Patent Application No. PCT/KR2017/002262 filed on Mar. 2, 2017, under 35 U.S.C. § 371, which claims priority to Korean Patent Application Nos. 10-2016-0088071 filed Jul. 12, 2016 and 10-2017-0018835 filed Feb. 10, 2017, respectively, which are all hereby incorporated by reference in their entirety. TECHNICAL FIELD The present invention relates to a PCL patch tissue regeneration scaffold and a method of preparing the same, and more particularly, to a polycaprolactone (PCL) patch tissue regeneration scaffold having a growth factor release type and a method of preparing the same. BACKGROUND ART Chronic otitis media (chronic tympanic perforation) is one of the most common causes of hearing damage in the otolaryngology area, in which the perforation of the tympanic membrane continues, however, most of the therapeutic methods still rely on surgical methods (tympanum regeneration). The perforation of the tympanic membrane can be largely divided into acute and chronic. Acute tympanic perforation is likely to be regenerated naturally if the perforation size is not large, however, most chronic tympanic perforation (chronic otitis media) of at least 3 months old, is not treated naturally, and as it gets older, it can induce complications such as otorrhea, pyopoiesis, hearing loss, facial nerve paralysis or brain abscess. To treating chronic otitis media, it is generally necessary to use a surgical method called tympanum regeneration, which is costly, requires complicated surgery for an operation time of 1 hour or more and anesthesia, hospitalization, and has a problem for recurrence rate of at least 10%. According to the National Statistical Office, from year 2009 to 2013, the number of patients with tympanitis is estimated to be about 1.7 million, and the cost of medical treatment has reported about 170 billion won. Many methods have been studied to overcome the problems of these surgical methods, the most classic of which is the paper patch technique. In other words, it is a treatment that allows marginal cells of tympanic perforation to use a paper patch as a support by cutting and pasting a paper patch on the perforation site. Generally, it is effective for small perforation of acute tympanic perforation, but it has no effect on chronic tympanic perforation. In other words, non-biocompatible materials cause inflammation and the success rate is less than 10%. In addition to the paper patch technique, during a patch-type scaffold for treating chronic tympanic perforation, chitosan patches containing growth factors was produced and then applied to a chronic animal model to confirm the therapeutic effect. Growth factors were supported on the chitosan patch so as to have a sustained release and it was confirmed that the cure rate was high in the animal model with chronic tympanic perforation. The epidermal growth factor (hereinafter, referred to as ‘EGF’) supported on the chitosan patch is known to be a factor affecting the proliferation of stem cells, and it has been reported that a patch supporting the same stimulates stem cells remaining in the margin of the tympanic perforation to affect chronic tympanic perforation treatment. However, since the chitosan patch has not been approved by the FDA of the US Food and Drug Administration, various attempts have not been made to easily apply it to clinical practice. All cells in the human body are in various environments provided by the extracellular matrix (ECM), and the extracellular matrix has been reported to have each specific nanostructures. Recent studies have shown that in vitro experiments can control cell morphology, mobility, and function by nanostructures of the substrate surface on which cells was adhered and grown. Thus, in the case of the tympanic tissue, it is suggested that the tympanic stem cell may change its shape, mobility and function in response to the shape or arrangement of the nanostructure, if the proper nano-environment is provided. Electrospinning is one of the easiest methods to produce nanofibers, which can produce nanofibers that are uniform, continuous and can exhibit various properties depending on the composition ratio. At this time, the portion in which the nanofibers spun by the electrospinning method are obtained is called a collector, and the characteristics of the nanofibers obtained according to the collector shape can be controlled. Through active research at home and abroad, we have found that cell characteristics are regulated in nanostructures obtained by electrospinning. However, there is no method for manufacturing a spindle-shaped tissue regeneration scaffold by electrospinning a biodegradable and biocompatible polymer that can be used for drug delivery and various medical field researches. DISCLOSURE Technical Problem