• 검색 결과가 없습니다.

Nanobiomaterials for stem cell and tissue engineering

N/A
N/A
Protected

Academic year: 2022

Share "Nanobiomaterials for stem cell and tissue engineering"

Copied!
41
0
0

로드 중.... (전체 텍스트 보기)

전체 글

(1)

Nanobiomaterials

for stem cell and tissue engineering

화학생물공학부 김병수 교수

(2)

Medical Approach

Pharmacol.

Management

Organ

Transplantation

Kidney, BM, Liver Heart, Pancreas, etc.

Regeneration of Organ

Stem Cell Therapy

& Tissue Engineering

미래의학 = 재생의학 (再生醫學) Regenerative medicine

(3)

Prof. S. Yamanaka (2012 Nobel Prize)

Human disease model

Drug screening

(4)

Research is presently being conducted on several different types of tissues and organs, including:

 Skin

 Cartilage

 Blood Vessels

 Bone

 Muscle

 Nerves

 Liver

 Kidney

 etc. etc. etc.

Tissue Engineering

is the

in vitro

development of tissues or organs to replace or support the function of defective or injured body parts, or the directed management of the repair of tissues within the body.

(5)

출처 : Regenerative medicine market, Trimark, 2013

(6)

Injection of MSCs to infarction area through catheter

(Intracoronary injection) Autologous

bone marrow aspirate

MSCs Infarction

area In vitro culture

Limitations :

5% increase in ejection fraction (heart function)

Likely due to

 Low possibility of stem cell differentiation into cardiomyocyte

 Wash-out of transplanted stem cells from the injection area

FCB-Pharmicell, Inc (Korea)

Approved clinical use of bone marrow stem cells

for myocardial infarction treatment

(7)

Cardiac stem cell sheets

for myocardial infarction treatment

Prof. Teruo Okano

(Tokyo Women’s Medical University)

(8)

Proarrhythmic (부정맥) potential of MSC transplantation to myocardial infarction heart

Due to no electric coupling of inexcitable MSCs to cardiomyocytes (심근세포)

(9)

Overexpression of connexin 43 in skeletal myoblasts

prevents ventricular arrhythmias following transplantation

S. Fernandes et al., J. Cell. Mol. Med. 13; 3703-3712, 2009

Connexin 43 overexpression in myoblasts improved intercellular electrical coupling between transplanted myoblasts and cardiomyocytes in infarcted heart

VT : ventricular tachycardia (심실성빈맥)

Ventricular hyperexcitability of the myocardium after myoblast transplantation

Connexin 43

(a gap junction protein)

control : myoblasts

null : myoblasts + empty vector

Cx43 : myoblasts + connexin43 gene

(10)

M. Shevach et al., J. Mater. Chem. B, 2013, 1, 5210 (A) Fabrication of PCL–gelatin fiber scaffolds by electrospinning. (B) Evaporation of gold NPs onto the fibers to create nanocomposite scaffolds. (C) Cardiac cells are seeded in the scaffolds for (D) engineering a functional cardiac tissue

To increase the electrical conductivity and enhance the electrical signal transfer between cardiac cells

Gold nanoparticle-incorporated cardiac patch

(11)

M. Shevach et al., J. Mater. Chem. B, 2013, 1, 5210

Cardiac sarcomeric actinin immunostaining

scaffolds without gold Gold NP scaffolds

Engineered cardiac tissue function

Normal crdiomyocytes

Gold nanoparticle-incorporated cardiac patch

(12)

Au - - + +

Electric

stimulation - + - +

Cx43 Intensity

J.O. You, et al., Nano Lett. 2011, 11, 3643–3648

Conductive scaffolds enhance connexin 43 expression

(13)

D.H. Kim et al., Integr. Biol., 2012, 4, 1019–1033

well-aligned myocardium

Nanopatterned cardiac patches promote myocardial

regeneration by cardiosphere-derived cells

(14)

D = 283 nm D = 749 nm

Christopherson GT, et al. Biomaterials (2009)

Control of neural stem cell differentiation

with nanofiber diameter

(15)

Nestin+ : progenitor Tuj1+ : neuron

RIP+ : oligodendrocyte GFAP+ : astrocyte

Fiber D = 749nm

283nm

Control of neural stem cell differentiation with nanofiber diameter

Christopherson GT, et al. Biomaterials (2009)

(16)

Human fetal tendon stem cell

seeding

Aligned Random

Nanofibers

CFDA

6 weeks in vivo

Yin Z, et al. Biomaterials (2010)

teno-lineage differentiation

Effects of nano-fiber alignment on cell

alignment and tendon differentiation

(17)

Within bone, type I collagen has parallel array of collagen fibrils 10 - 300 nm in size.

Pitch = 400 nm 1400 nm 4000 nm planar control

Nanotopographical pattern modulates osteogenic differentiation of MSCs

17 Watari S, et al. Biomaterials (2012)

(18)

B.K.K. Teo et al., ACS Nano 2013, 7: 4785

Nanotopography modulates focal adhesion and

differentiation of stem cells

(19)

# of focal adhesion per cell

3D AFM height images of Ti surfaces

8 nm 15 nm

MSCs stained for actin and osteocalcin

planar Ti 8 nm 15 nm nanodots

T. Sjöström et al., Adv. Healthcare Mater. 2013, 2, 1285–1293

Nanopatterning of titanium implants for enhancing

osteointegration

(20)

Topographically defined implant for enhanced osteoconduction

DAPI F-actin

Guided osteoblast migration in vitro

Tissue Engineering 22: 654-664 (2016) Implants with different topography

Width of pits = 5µm

(21)

Enhanced bone regeneration by topographically defined implant

Topographed implant (diameter = 5 mm)

calvarial defect (diameter = 4 mm)

Regenerated bone tissue

7 weeks Osteoblast

migration

Mouse calvaria

Implant fixation with fibrin gel

Tissue Engineering 22: 654-664 (2016)

(22)

 FDA approved for spine fusion, bone fracture, and dental bone grafting

 InFUSE® (USA)/ InductOS ® (Europe) : BMP-2 soaked in bovine type I

collagen sponge

Source: www.infusebonegraft.com

 OP-1 ® : BMP-7 mixed with bovine bone collagen and sterile saline solution to form a paste

Clinical use of BMP-2 and -7

(23)

Sustained release of BMP-2 for effective bone regeneration

(24)

OH

O

O

O

C O OH C C

O O

O O O

O O

OH

O C O

HO O

OH OH OH OH

OH O

O

O

OO OO O

O

O • Hydrophobic amino acids (A, F, G, I, L, M, P, V, W, Y)

Positively charged amino acids (K, R) Bone Morphogenetic Protein 2

Hydrophobic π domain Negatively charged

oxygenated group

Graphen Oxide Surface

“Hydrophobic Interaction”

“Electrostatic Interaction”

Ti/GO- (Outer-most Layer: GO-)

GO-COO- GO-NH3+

Ti

GO-COO- (GO-) GO-NH3+ (GO+)

Layer-by-Layer (LbL) Assembly

Delivery of BMP-2 protein for osteointegration of dental implants

La WG et al., Small 2013, 9(23): 4051-4060

(25)

0 2 4 6 8 10 12 14 16 0

20 40 60 80 100

Ti Ti/GO+

Ti/GO-

Cumulative release (%)

Time (Days)

*

*

*

*

Release of BMP-2

Lower Loading Amount

& Higher Initial Burst

Sustained Release Ti

Ti/GO-

With BMP-2 Without BMP-2

Ti or Ti/GO- rings with or without BMP-2

Screws

La WG et al., Small 2013, 9(23): 4051-4060

Delivery of BMP-2 protein for osteointegration of

dental implants

(26)

Microfracture ACT Matrix assisted ACT

Current techniques for cartilage repair

E.A. Makris et. al. Nature Reviews 2014

(27)

Strengths

Regeneration of hyaline-like cartilage

Limitations

Multiple surgery procedures

Expensive cost due to in vitro cell expansion step

Patients’ waiting during the in vitro cell expansion period

Dedifferentiation of chondrocytes occurs during in vitro culture

Invasive surgery

Autologous chondrocyte transplantation (ACT)

(세원셀론텍)

(28)

ACT using scaffold (Bioseed-C, 독일 Biotissue 사)

• Autogenous chondrocytes seeded on PLGA-poly-p- dioxanone scaffold

• Advantages over ACT :

- No harvesting of periosteum

- Arthroscopical implantation possible ; less invasive

• Limitations - 2번 수술

- 세포배양하므로 가격 비쌈

3x2x0.2cm

9 months later

(29)

Transplantation of cells harboring therapeutic gene (코오롱생명과학, 미국에서 임상 3상 시험)

TGF-ß Gene

치료 유전자 (TGF-beta)를 바이러스벡터를 이용하여

타가 연골세포에 삽입

Cell + therapeutic gene 관절내 이식

(30)

1 주후 6 주후

TGF-β1

Control

Transplantation of cells harboring therapeutic

gene (토끼 동물시험)

(31)

돼지연골세포를 배양한 후 세포를 제거하고

남은 세포외기질

Biofilm to enhance marrow stimulation- mediated cartilage regeneration

㈜리젠프라임, ArtiFilm

TM

줄기세포의 연골분화 유도 안됨

→ 섬유연골 재생 가능성

(32)

Scaffold to enhance marrow stimulation-mediated cartilage regeneration

(미국 Cartilix사, ChonDuxTM)

(33)

3D printing of organ

• Cell spheres are fused to form vessels

• Advantages

- Precise control of robotic bio- fabrication of organs and vessels - Fabrication of patient-specific

shaped organs

www.explainingthefuture.com

(34)

생체고분자 틀 INJECTION

3D bioprinting

(35)

• 3D프린팅 기술로 만든 기도 부목이 생후 2개월이었던 아기의 생명을 구했다.

• 이 아기는 숨을 쉬는 동안에도 기도가 약해서 기도벽이 무너져 내리는 증상 을 가졌다.

• 기도 부목은 3년 동안 아기의 기도가 정상적으로 자라나도록 도운 뒤 자연스

럽게 몸 속에서 생분해 됐다.

3D bioprinting of airway splint (기도 부목)

N Engl J Med 2013; 368:2043-2045

(36)

3D bioprinting of airway splint

A : the airway before placement of the splint

B : the patient-specific computed tomography–based design of the splint (red)

C : an image-based 3D printed cast of the patient's airway without the splint in place D : the splint in place

E : intraoperative placement of the splint (green arrow) F : the airway after placement of the splint

G : the airway 1 year after placement of the splint

N Engl J Med 2013; 368:2043-2045

(37)

얼굴뼈 종양을 제거한 후, 눈 선상이 일직선이지 않은 상태의 CT를 찍어 임플란트를 디자인하고 이를 3D 프린터로 출력하여 생분해성 임플란트를 제작함. 이를 이식하고 2주 경과 후 모습.

http://ims.postech.ac.kr/

3D bioprinting of bone

(38)

3D 프린팅 기술로 맞춤형 치아 교정기와 보청기 제조

(39)

3D bioprinting of absorbable stent

S.A Park etal. Materials Letters141(2015)355–358

(40)

 신약 개발에 있어서, 동물실험에서 확인하기 어려운 부분을 인체유래 세포에서 생성된 인공 조직이나 인공기관으로 대체할 수 있음

 유럽은 동물실험을 이용한 화장품 및 화장품원료 판매금지

 피부와 관련된 독성시험이 동물실험 대신 세포실험 등으로 표준화되는 추세

인공피부를 이용한 피부자극성 및 독성 실험

동물대체시험법으로서 인공조직 및 인공기관모델 개발

(41)

동물대체시험법으로서 인공조직 및 인공기관모델 개발

표경호 BRIC View 2015-T07

참조

관련 문서

Growth and rosmarinic acid accumulation in callus, cell suspension, and root cultures of wild Salvia fruticosa.. Plant Cell Tissue

Meterials and Methods: Our study involved three stem cell sources-subacromial bursal tissue, bone marrow, and umbilical cord-derived stem cells. Under consent we

Recent stem cell studies have reported that cultured hematopoietic stem cells (HSCs) are reactivated through fetal hemoglobin expression by treatment with

– Embryonic stem cell : Pluripotent cells that can give rise to all tissue types. – Adult stem cell : Multipotent cells

The porous PCL scaffolds produced from the gas foaming and salt leaching and plasma surface treatment are suitable for potential applications in bone

Therefore, cTnT is a recommended biomarker for use in the detection of myocardial infarction (MI) and in acute coronary syndromes.[8] Indeed, several authors

Integrative STEM(Science, Technology, Engineering, and Mathmatics) education: Contemporary trends and issues.. The concept of creativity: Prospects

미국에서는 최초로 Virginia Tech에서 처음으로 대학원 기술교육 전공과정에 통합적 STEM 교 육 전공과정(Integrative STEM)을 개설하면서 본격적으로 프로그램