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276 B. N. H. M. Neeroa et al.
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Hosseinifar
Low et al.
Increased apoptotic-mediated cell
(2019)
death
Ektate et al.
(2018)
Enhanced tumour-associated M1
phenotype macrophage activities
Moghimipour
et al. (2018)
Enhanced cytotoxicity activity of
5-FU
Serini et al.
(2018)
Shen et al.
doxorubicin
Prevented systemic absorption and
(2019)
improved drug accumulation at the
target site
Wei et al.
(2020)
Improved cellular uptake in TfR
overexpressing cells and increased
tumour accumulation and
inhibition
Ramezani
et al. (2020)
Improved tumour growth
inhibition by nucleolin-targeting
and drug-controlled release by
MMP-2 protein
et al. (2018)
release via polymer degradation,
improved cellular uptake and
Enhanced target-specific drug
promote apoptosis
t:1 Table 1 Effect of different types of NPs with their associated therapeutic response on colorectal cancer
Type of
nanosystem Description Anticancer agent Model Therapeutic effect References
t:2
mice
cancer cell line
Curcumin In vitro HCT116 colon
Thermo-sensitive liposomes Doxorubicin In vivo BALB/c female
Magnetic cellulose nanocrystals
(MCNC)-PE
Lipid-
based NPs
t:3
t:4
cancer cell line
Folic acid liposomes 5-Fluorouracil In vitro CT26 colon
t:5
In vivo BALB/c male
mice
t:6
In vivo CT26 tumour-
bearing BALB/c mice
cancer cell line
Doxorubicin In vitro HCT116
Resveratrol-stearate SLNs Doxorubicin In vitro HT29 CRC line Enhanced antineoplastic activity of
Dextran-folic acid SLNs Doxorubicin In vitro CT26 colon
Tf-binding peptide-
Polymer-
t:7
t:8
t:9
In vivo BALB/c female
cells
mice
human colon cancer
functionalised polymersomes
based NPs
t:10
t:11
In vivo C26
subcutaneous tumour
In vitro C26 mouse
colon cancer cells
7-ethyl-10-hydroxy
camptothecin
AS1411 aptamer conjugated,
PEG-peptide-PLA
t:12
model BALB/c mice
(SN38)
polymersomes
5-Fluorouracil In vitro HT29 colon
β-Cyclodextrin-alginate (β-CD,
t:13
cancer cells
AL) nanogels
t:14
Development of Biocompatible Nanocarriers for the Treatment of Colorectal Cancer 277
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(continued)
Shad et al.
(2020)
release, increased drug half-life
Enhanced target-specific drug
Ramzy et al.
(2020)
and promote apoptosis
Improved drug solubility from
encapsulation, increased
cytotoxicity on sigma receptor
overexpressing in HT29 cells
Reimondez-
Troitiño et al.
(2019)
Reduced cell migration and
proliferation with increased
cancer-killing activity in curcumin
and improved cellular uptake of
Priyadarshi
et al. (2021)
miR-145
Improved controlled release of
drugs, promoted inhibition of cell
proliferation and migration,
cellular uptakes, apoptotic
signalling and release of
Pishavar et al.
(2019)
pro-inflammatory cytokines in
HCT116 cells
Increased cytotoxicity of drug and
inhibit tumour growth rate
Näkki et al.
(2019)
Comparable cytotoxicity activity
with free drugs but exhibited
reduced side effects
cancer cells
Oxaliplatin In vitro HT29 colon
Hyaluronic acid coated-folic acid
functionalised (HA-FA) alginate
t:15
Thymoquinone In vitro HT29, HCT116
nanogels
Anisamide-eudragit S100
and Caco-2 cells
polymeric nanocapsule
t:16
SW480 human CRC
Protamine nanocapsules Curcumin and
cells
miR-145
t:17
carcinoma and HT29
CRC adenocarcinoma
Gallic acid In vitro HCT116 CRC
Poly(amidoamine) (PAMAM)
dendrimers
t:18
In vivo C26 BALB/c
In vitro C26 mouse
Doxorubicin and
Alkyl-PEG-cholesteryl
male mice
colon cancer cells
TRAIL plasmid
chloroforate-modified PAMAM
dendrimers
t:19
t:20
In vivo CT26
cell line
Doxorubicin In vitro CT26 cancer
-PEG)
3
Superparamagnetic iron oxide/
calcium carbonate/polyethylene
glycol (SPION-CaCO
Inorganic
NPs
t:21
subcutaneous tumour-
bearing BALB/c nude
mesoporous silica
mice
t:22
278 B. N. H. M. Neeroa et al.
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Babaei et al.
Hao et al.
Promoted caspase-dependent cell
(2019)
death and apoptosis from
mitochondrial damage and
autophagy
Khan et al.
(2020)
Enhanced inhibition of cell
proliferation and promoted
Lo et al.
(2020)
apoptosis
Increased NP cellular uptake,
enhanced tumour growth
suppression
Shabbir et al.
(2021)
Increased HCT116 cell sensitivity
towards drug’s growth inhibitory
effect, preventing tumour
(2020)
release, apoptosis initiation and
formation
Improved drug’s controlled
in vivo anti-tumour effect.
Increased cellular uptake via call-
targeting AS1411 DNA aptamer
Prajapati et al.
(2019)
release, enhanced inhibition of cell
Improved drug’s controlled
proliferation due to internalisation
by receptor-mediated endocytosis,
increased cytotoxicity
Type of
nanosystem Description Anticancer agent Model Therapeutic effect References
Table 1 (continued)
t:24
In vivo HCT116
cell lines
therapy
silver (PD/Au-Ag) metal NPs
t:23
subcutaneous tumour-
In vitro HCT116 cancer
Photothermal
Polydopamine-coated gold-
bearing BALB/c nude
mice
t:24
cell lines
Curcumin In vitro HCT116 cancer
Eudragit RS 100 modified
quantum dot NPs
t:25
In vivo HCT116
In vitro HCT116 cancer
cell lines
Doxorubicin and
EFGR (epidermal
growth factor
PEI-folic acid (PIF) graphene
quantum dot NPs
t:26
subcutaneous tumour-
bearing BALB/c nude
receptor)
mice
177Lu-labelled Au NPs Cetuximab In vitro HCT116,
t:27
HCT68 and SW620
colon cancer cell lines
t:28
In vivo C26
colon cancer cells
Camptothecin In vitro C26 mouse
PEG-survivin shRNA
expressing plasmid (iSur-pDNA)
tagged with AS1411 DNA
t:29
subcutaneously
tumour-bearing female
BALB/c mice
Gemcitabine In vitro HT29 colon
aptamer
Mesoporous silica
Hyaluronic acid-PEG (HA-PEG)
t:30
In vivo Sprague-
cell line
conjugated multi-walled carbon
nanotubes
t:31
Dawley rat
t:32
Development of Biocompatible Nanocarriers for the Treatment of Colorectal Cancer 279
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and –OH functional groups, resulting in various possibilities of chitosan
2
2493 Construction of Biocompatible Nanocarriers for Colorectal
250Cancer Treatment
2513.1 Biomaterials Used for Biocompatible Drug Nanocarrier
252Synthesis
253Various biomaterials can be used in the generation of drug nanocarriers. This review
254will focus on the most promising ones in the subfield of colorectal cancer treatment,
255namely chitosan, xanthan gum, cellulose and alginate. Chitosan is a natural polysac-
256charide derived from chitin and is made by deacetylation to form d-glucosamine
257and N-acetyl-d-glucosamine linked by 1-4-β-glycosidic bonds (Ways et al. 2018).
258Chitosan has mucoadhesive properties which makes it suitable for drug release in
259various epithelial systems including inte stinal, nasal, eye and pulmonary. Referring
260to the structure, chitosan can form covalent and hydrogen bonds due to the presence
261of –NH
262derivatives. These functional groups are also essential in determining the solubility
263of chitosan. Under low pH, the chitosan becomes positively charged as its amine
264groups are protonated. The positively charged amino groups from the polymer can
265then form hydrogen bonds and strong ionic interactions with the negati vely charged
266epithelial surface of the mucus (Ways et al. 2018). For example, a chitosan lactate
267gel was successfully developed and patented for the controlled release of lactic acid
268onto the vaginal mucosa for vaginal maintenance therapy (Sandri et al. 2012). In
269another study conducted by Cerchiara’s group (2015), they found chitosan
270synthesised using spray-drying method was an effective nanocarrier for vancomycin
271in treating serious colorectal inflammation. Thus, chitosan is a promising candidate
272to act as a nanocarrier for the treatment of colorectal cancer.
273Xanthan gum is a high molecular weight heteropolysaccharide produced
274by Xanthomonas campestris. This material is a polyanionic polysaccharide, with
275pentasaccharide subunits, and glucuronic acid and pyruvic acid side chains (Malik
276et al. 2020). It has been extensively used for synthesising nanocarriers due to its
277outstanding biochemical properties, exhibiting excellent biocompatibility, water
278solubility and maintaining high functionality under astringent environments such
279as high salt or acid (Kang et al. 2019). Xanthan gum has been developed to act as a
280drug-controlled released matrix polymer for oral administered product s. In a study,
281they developed aceclofenac tablets coated with xanthan gum for colon-targeted
282therapy. They found that the tablets were able to resist drug release in the gastroin-
283testinal tract but could dissolve very well in the colon. It was also noted that the drug
284release was more efficient under a weak acidic/neutral pH rather than a highly acidic
285environment, which makes it a suitable colon-targeting agent (Singhvi et al. 2019).
286Cellulose is the most abundant natural polymer found in nature; it is made up of
287β-1-4 linked glucopyranose unit, forming a high-molecular-weight linear homopol-
288ymer. They are highly polymorphic, and their properties may differ due to the variety
289of sources and different production/extraction methods to synthesise cellulose
290(Lugoloobi et al. 2021). They are widely used in drug delivery systems mainly
291due to their abundance and low cost in manufacturing alongside their high degree of
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292 flexibility which allows for modification in their solubility and gelation (Lugoloobi
293 et al. 2021). Cellulose is often functionalised to form cellulose ethers or cellulose
294 esters as they have poor solubility due to their intermolecular hydrogen bonding.
295 Cellulose ether derivatives are commonly favoured as nanocarriers for oral dosages
296 due to their swelling capacity, which allows the hydrogel layers to control the drug
297 diffusion and delivery (Amalin Kavitha et al., 2020). Recently, Li et al. (2019)
298 successfully synthesised cellulose nanofiber as a nanocarrier for quercetin, an
299 antioxidant, which showed a higher delivery profile than control. Cellulose
300 nanocrystals are also synthesised which offer additional advantages compared to
301 the original nanocarriers (Yusefi and Shameli 2021). In anothe r study, cellulose
302 nanocrystals were able to encapsulate curcumin for antimicrobial application and
303 improve the stability of the formula at different pHs (Asabuwa Ngwabebhoh et al.
304 2018).
305 Another biopolymeric material that has been widely used in drug delivery is
306 alginate. Alginates are negatively charged salts that came from alginic acid, com-
307 posed of 1,4-linked β-D-mannuronic acid (M) with α-L-glucuronic acid (G) that are
308 formed in an irregular block-wise pattern (He et al. 2020; Osorio et al. 2020). Similar
309 to chitosan, alginate has excellent mucoadhesive properties which allow its use
310 as nanocarriers in various epithelial cancers. Sookkasem et al. (2015) formulated
311 alginate beads, on a macro scale, which could encapsulate curcumin and allow colon
312 therapy to be more target specific. The alginate beads prevented the rapid physiolog-
313 ical clearance of curcumin in the upper gastrointestinal tract and were only released
314 in the colon. In another study in 2018, Freitas and team designed mucoadhesive
315 sericin-alginate particles which were thought to improve sustained drug release
316 possibly via a dissolution mechanism (Freitas et al. 2018).
317 3.2 Potent Nanocarriers Against Colorectal Cancer
318 This section focuses on the current update of nanocarriers that show potent killing
319 against colorectal cancer. Over the years, studies have shown that colon cancer cells
320 overexpress several proteins and cancer-specific molecules which make them as
321 ideal therapeutic targets. These include CD98 (Xiao et al. 2018), CD44 (Jia ng et al.
322 2018), galectins (Liu et al. 2018), biotin receptors (Lin et al. 2018) and microRNAs
323 (miRNAs) (Zheng et al. 2018). Recent development of nanocarriers was also
324 designed to target these molecules. On top of that, FDA-approved anticancer drugs
325 such as 5-FU, irinotecan, oxaliplatin, doxorubicin, bevacizumab and capecitabine as
326 well as other cancer-killing compounds such as curcumin, silymar in and colchicine
327 were also either encapsulated or conjugated with the nanocarriers to improve the
328 treatment efficacy.
329 CD98 receptors overexpressed on the apical membranes of colon cancer cells,
330 Xiao et al. (2018) synthesised a CD98-siRNA and camptothecin-loaded PEGylated
331 Fab-NPs embedded in a hydrogel for colon targeting using mouse models. The
332 efficacy of combination drug was higher than the nanoparticles containing a single
333 drug due to higher drug internalisation into the tumour cells. In another study, Jiang
Development of Biocompatible Nanocarriers for the Treatment of Colorectal Cancer 281
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334et al. (2018) prepared hyaluronidase (HA)-conjugated mesoporous silica NPs loaded
335with 5FU and demonstrated their cytotoxicity to colon cancer cells. HA on the
336surface of NPs targeted the CD44 receptors overexpressed in the cancer cell s.
337Similarly, galectins are overexpressed and found to play crucial modulatory roles
338in colon cancer. Liu et al. (2018) developed 5FU-loaded mesoporous silica
339NP-based galactosylated chitosans for colon cancer-specific drug delivery and
340showed potent killing against the cancer cells compared to the free 5FU in vitro.
341On the other hand, Lin et al. (2018) developed poly (ethylene glycol) and biotin-
342modified Dox-loaded silica NPs which target biotin receptors. The tumour cells were
343effectively killed in both HCT116 colorectal cancer cell line and tumour-bearing
344mice. In addition to cell receptors, some miRNAs are also found to regulate cancer
345events. Zheng et al. (2018) synthesised poly (D,L-lactide-co-glycolide)/poly
346(L-lactide)-block-poly(ethylene glycol)-folate polymer NPs that were loaded with
347miR-204-5p and demonstrated their anticancer effects on colon cancer cells and
348xenograft colon tumour models in vivo.
349Curcumin, a substance in turmeric plants, has exhibited various pharmacological
350properties such as antimicrobial (Teow et al. 2016) and anticancer actions (Tomeh
351et al. 2019). To further enhance the anticancer activity against colon cancer cells,
352curcumin has been encapsulated into various types of nanocarriers. For instance,
353Alkhader et al. (2018) synthesised a chitosan-pectinate NP system (CUR-CS-PEC-
354NPs) which could enhance the oral bioavailability of curcumin by protecting it from
355gastric degradation. A pH-responsive xylan-curcumin prodrug NPs could release the
356loaded curcumin at acidic pH and improve the anticancer killing against colon
357cancer cells compared to curcumin alone (Sauraj Kumar et al. 2018). Interestingly,
358codelivery of curcumin and 5-FU from a xylan-SS-curcumin redox-sensitive
359prodrug NPs could effectively inhibit colon cancer cells (Kumar et al. 2020). The
360same research group also constructed xylan-5-FU-1-acetic acid conjugates (Kumar
361et al. 2017) and amphiphilic xylan-stearic acid-based NPs (Kumar et al. 2019) which
362improved the anticancer activity of 5-FU against colon cancer cells upon delivery.
363AbouAitah et al. (2020) generated a mesoporous silica NPs functionalised with
364phosphonate groups and folic acid chitosan-glycine complex to deliver colchicine
365against colon cancer cells. Colchicine targeted and inhibited the cancer cells by
366inducing apoptosis. In another study, an epidermal growth factor- (EGF-)
367functionalised poly(lactic-co-glycolic acid) (PLGA) NPs successfully deliver 5-FU
368and perfluorocarbon for effective treatment of colon cancer. The functionalised NPs
369were able to selectively recognise specific receptors present on colon cancer cells
370and inhibit tumour growth (Wu et al. 2020). Marcelo et al. (2020) used mesoporous
371NPs encapsulating fluorescent silica quantum dots (SiQDs) to act as cargo for
372doxorubicin in killing colon cancer cell line, HCT116 and HT29 cells. They found
373that MSNP-doxorubicin showed the most significant cytotoxic effect in both cell
374lines, with a higher uptake against HCT116, while free drug doxorubicin exhibited
375the lowest uptake. This confirms that doxorubicin-loaded MSNPs can elicit signifi-
376cant cancer-killing properties. On the other hand, the nanocarrier with the lumines-
377cent SiQDs (SiQD-MSNP-doxorubicin) was postulated to undergo cellular
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378 internalisation but resisted immediate drug release of doxorubicin, showing
379 possibilities of controlled drug release of doxorubicin (t > 72 h).
380 Using an oxaliplatin-loaded PLGA microsphere, Lagarce et al. (2002) showed
381 high entrapment efficiency (up to 90%) and diverse release profiles, which collec-
382 tively improved the efficiency of local tumour treatmen t. Liu et al. (2015) loaded
383 5-FU onto a hyaluronic acid (HA)-conjugated silica nanocarriers for colon cancer
384 treatment. The nanocarrier could attach and accumulate in colon cancer cells due to
385 their HA surface alterations, this resulted in enhanced antitumour action and
386 decreased side effects as compared to non-modified nanocarriers. Lee et al. (2006)
387 generated docetaxel-loaded biodegradable dendrimers with multiple attachment sites
388 capable of releasing drug in a controlled manner. These drug-loaded dendrimers
389 were ten times less toxic than free docetaxel in the colon cancer treatment.
390 4 Limitations and Future Perspectives
391 Over the last few decades, nanomedicine has emerged and amassed tremendous
392 growth in its potential to revolutionise cancer therapy. Massive investments, in terms
393 of finance and labour, have been exhausted to design, research and develop NPs in
394 hopes of implementing nanomedicine into the healthcare system. However, very few
395 made it to the clinical trial phase and even fewer were approved for clinical use
396 (Desai 2012). In this section, the current challenges and limitations of NPs develop-
397 ment are discussed.
398 To succeed in the clinical translation of nanoparticle-based therapeutics, an
399 understanding of the biodistribution, pharmacokinetics, safety mechanisms, and
400 mechanism of action of NPs must be full y establis hed. However, the long-term
401 inflammation, toxicity and carcinogenesis of these NPs are not fully understood yet
402 (Desai 2012). This uncertainty possesses one of the biggest hurdles for nanoparticle-
403 based drug carriers to progress onto clinical trials. Without having a clear under-
404 standing on the route of NP delivery and outcomes, patients undergoing this therapy
405 would have increased the risk of developing unforeseen adverse reactions. Hence, it
406 is necessary to develop in vitro and animal models tailored to investigate the main
407 goal of NP-based drug-delivery systems which is to maximise the potency and
408 efficacy of the therapy whi le reducing systemic side effects (Park 2013). In order
409 to achieve this objective, it is crucial to use these models to (i) study about the
410 interaction between NPs and cells, (ii) optimise the transcytosis and uptake rate of
411 NP into the tumour microenvironment, (iii) design NPs that are able to overcome the
412 biological barriers present in the body such as the reticuloendothelial system and
413 renal system and (iv) determine the clearance rate and degradation of nanocarriers
414 (Park 2013; de SL Oliveira et al. 2021). Essentially, in-depth studies on these grey
415 areas will ensure that future nanoparticle-based drug carriers would have a smooth
416 transition from preclinical studies to clinical settings and subsequently be validated
417 for clinical use.
418 Another limitation of existing nanoparticle-based drug carriers is their delivery
419 efficacy and bioavailability which severely limits their therapeutic potency. To
Development of Biocompatible Nanocarriers for the Treatment of Colorectal Cancer 283
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420increase the delivery efficiency of nanoparticle-based therapeutics to the tumour site,
421the properties of NPs must be exploited or modified to extend circulation in blood
422(bioavailability) and minimise non-specific clearance rate by phagocytosis and the
423liver (Sabit et al. 2022). For instance, it was found that non-spherical NPs have
424higher bioavailability and are less vulnerable to mononuclear phagocyte system
425(MPS) sequestration in blood vessels compared to their counte rpart (Ye et al.
4262018). At the same time, functionalisation of NPs with active ligands specificto
427tumours will ensure maximum accumulation in tumour microenvironment compared
428to other sites which further limits free drug toxicity (Manzoor et al. 2012). Moreover,
429functionalisation NPs are shown to have decreased serum protein absorption and are
430resistant to macrophages which also enhances their bioavailability (Ye et al. 2018).
431Hence, further research and modifications should be done on existing nanoparticle-
432based drug carrier to develop drug carriers that protect the cargo (i.e., drugs) from
433degradation, phagocytic activities and non-specific clearance. By having long bio-
434availability, lower drug dosage is required. This will reduce the incidence and
435severity of side effects, and lower cost of treatment, contributing to better treatment
436outcome and high quality of life post-treatment.
4375 Conclusion
438The use of biocompatible drug nanocarrier is a promising treatment method for
439colorectal cancer due to its ability to effectively penetrate the targeted tumour area
440and delivery anticancer drugs without compromising normal cells. NPs have rich
441surface chemistry and flexibility in size and shape modifications which makes them
442ideal drug carriers to overcome the limitations of conventional drug delivery
443methods. Despite having many challenges in understanding the mechanism of
444actions and the delivery methods of NPs, it is crucial to push forward with the
445development of drug nanocarriers which maximises the potency of anticancer drugs
446in selectively killing colorectal cancer cells. Future studies should be done to
447elucidate the mechanisms of NPs and the long-term effects of NP usage in in vivo
448models and clinical trials to ascertain the safety and efficacy of nanoparticle-based
449therapeutics for clinical use. Nevertheless, as nanomedicine is a relatively new field,
450caution must be observed in conducting clinical studies while taking into account
451patients’ safety and ethics. With continuous development in nanomedicine, it is with
452great hope that advances in drug nanocarriers will improve the life expectancy and
453quality of life of colorectal patients, and cancer patients as a whole.
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